Peptides and compounds that bind to a thrombopoietin receptor

ABSTRACT

Receptors are peptides and peptide mimetics that bind to and activate the thrombopoietin receptor. Such peptides and peptide mimetics are useful in methods for treating hematological disorders and particularly, thrombocytopenia resulting from chemotherapy, radiation therapy, or bone marrow transfusions as well as in diagnostic methods employing labeled peptides and peptide mimetics.

This application is filed pursuant to 35 U.S.C. §371 as a United StatesNational Phase Application of International Application No.PCT/GB96/09623 filed Jun. 7, 1996 which claims priority from U.S. Ser.No. 08/485,301 filed Jun. 7, 1995 and now abandoned and U.S. Ser. No.08/478,128 filed Jun. 7, 1995 and now abandoned.

BACKGROUND OF THE INVENTION

The present invention provides peptides and compounds that bind to andactivate the thrombopoietin receptor (c-mpl or TPO-R) or otherwise actas a TPO agonist. The invention has application in the fields ofbiochemistry and medicinal chemistry and particularly provides TPOagonists for use in the treatment of human disease.

Megakaryocytes are bone marrow-derived cells, which are responsible forproducing circulating blood platelets. Although comprising <0.25% of thebone marrow cells in most species, they have >10 times the volume oftypical marrow cells. See Kuter et. al. Proc. Natl. Acad. Sci. USA91:11104-11108 (1994). Megakaryocytes undergo a process known asendomitosis whereby they replicate their nuclei but fail to undergo celldivision and thereby give rise to polyploid cells. In response to adecreased platelet count, the endomitotic rate increases, higher ploidymegakaryocytes are formed, and the number of megakaryocytes may increaseup to 3-fold. See Harker J. Clin. Invest. 47:458-465 (1968). Incontrast, in response to an elevated platelet count, the endomitoticrate decreases, lower ploidy megakaryocytes are formed, and the numberof megakaryocytes may decrease by 50%.

The exact physiological feedback mechanism by which the mass ofcirculating platelets regulates the endomitotic rate and number of bonemarrow megakaryocytes is not known. The circulating thrombopoieticfactor involved in mediating this feedback loop is now thought to bethrombopoietin (TPO). More specifically, TPO has been shown to be themain humoral regulator in situations involving thrombocytopenia. See,e.g., Metcalf Nature 369:519-520 (1994). TPO has been shown in severalstudies to increase platelet counts, increase platelet size, andincrease isotope incorporation into platelets of recipient animals.Specifically, TPO is thought to affect megakaryocytopoiesis in severalways: (1) it produces increases in megakaryocyte size and number; (2) itproduces an increase in DNA content, in the form of polyploidy, inmegakaryocytes; (3) it increases megakaryocyte endomitosis; (4) itproduces increased maturation of megakaryocytes; and (5) it produces anincrease in the percentage of precursor cells, in the form of smallacetylcholinesterase-positive cells, in the bone marrow.

Because platelets (thrombocytes) are necessary for blood clotting andwhen their numbers are very low a patient is at serious risk of deathfrom catastrophic hemorrhage, TPO has potential useful application inboth the diagnosis and the treatment of various hematological disorders,for example, diseases primarily due to platelet defects. Ongoingclinical trials with TPO have indicated that TPO can be administeredsafely to patients, in addition, recent studies have provided a basisfor the projection of efficacy of TPO therapy in the treatment ofthrombocytopenia, and particularly thrombocytopenia resulting fromchemotherapy, radiation therapy, or bone marrow transplantation astreatment for cancer or lymphoma. See, e.g., McDonald (1992) Am. J. Ped.Hematology/Oncology 14:8-21 (1992).

The gene encoding TPO has been cloned and characterized. See Kuter etal. Proc. Natl. Acad. Sci. USA 91:11104-11108 (1994); Barley et al. Cell77:1117-1124 (1994); Kaushansky et al. Nature 369:568-571 (1994);Wendling et al. Nature 369:571-574 (1994); and Sauvage et al. Nature369:533-538 (1994). Thrombopoietin is a glycoprotein with at least twoforms, with apparent molecular masses of 25 kDa and 31 kDa, with acommon N-terminal amino acid sequence. See, Bartley et al. Cell77:1117-1124 (1994). Thrombopoietin appears to have two distinct regionsseparated by a potential Arg-Arg cleavage site. The amino-terminalregion is highly conserved in man and mouse, and has some homology witherythropoietin and interferon-a and interferon-b. The carboxy-terminalregion shows wide species divergence.

The DNA sequences and encoded peptide sequences for human TPO-R (alsoknown as c-mpl) have been described. See Vigon et al. Proc. Natl. Acad.Sci. USA 89:5640-5644 (1992). TPO-R is a member of the haematopoietingrowth factor receptor family, a family characterized by a commonstructural design of the extracellular domain, including four conservedC residues in the N-terminal portion and a WSXWS motif (SEQ ID NO:1)close to the transmembrane region. See Bazan Proc. Natl. Acad. Sci. USA87:6934-6938 (1990). Evidence that this receptor plays a functional rolein hematopoiesis includes observations that its expression is restrictedto spleen, bone marrow, or fetal liver in mice (see Souyri et al. Cell63:1137-1147 (1990)) and to megakaryocytes, platelets, and CD34⁺ cellsin humans (see Methia et al. Blood 82:1395-1401 (1993)). Furthermore,exposure of CD34⁺ cells to synthetic oligonucleotides antisense to mplRNA significantly inhibits the appearance of megakaryocyte colonieswithout affecting erythroid or myeloid colony formation. Some workerspostulate that the receptor functions as a homodimer, similar to thesituation with the receptors for G-CSF and erythropoietin.

The availability of cloned genes for TPO-R facilitates the search foragonists of this important receptor. The availability of the recombinantreceptor protein allows the study of receptor-ligand interaction in avariety of random and semi-random peptide diversity generation systems.These systems include the "peptides on plasmids" system described inU.S. Pat. Nos. 5,270,170 and 5,338,665; the "peptides on phage" systemdescribed in U.S. patent application Ser. No. 07/718,577, filed Jun. 20,1991, U.S. patent application Ser. No. 07/541,108, filed Jun. 20, 1990,and in Cwirla et al., Proc. Natl. Acad. Sci. USA 87:6378-6382 (1990);the "polysome" system described in U.S. patent application Ser. No.08/300,262, filed Sep. 2, 1994, which is a continuation-in-partapplication based on U.S. patent application Ser. No. 08/144,775, filedOct. 29, 1993 and PCT WO 95/11992; the "encoded synthetic library"system described in U.S. patent application Ser. Nos. 08/146,886, filedNov. 12, 1993, 07/946,239, filed Sep. 16, 1992, and 07/762,522, filedSep. 18, 1991; and the "very large scale immobilized polymer synthesis"system described in U.S. Pat. No. 5,143,854; PCT Patent Publication No.90/15070, published Dec. 13, 1990; U.S. patent application Ser. No.07/624,120, filed Dec. 6, 1990; Fodor et al. Science 251:767-773(2/1991); Dower and Fodor Ann. Rep. Med. Chem. 26:271-180 (1991); andU.S. patent application Ser. No. 07/805,727, filed Dec. 6, 1991; each ofthe foregoing patent applications and publications is incorporatedherein by reference.

The slow recovery of platelet levels in patients suffering fromthrombocytopenia is a serious problem, and has lent urgency to thesearch for a blood growth factor agonist able to accelerate plateletregeneration. The present invention provides such an agonist.

SUMMARY OF THE INVENTION

This invention is directed, in part, to the novel and unexpecteddiscovery that defined low molecular weight peptides and peptidemimetics have strong binding properties to the TPO-R and can activatethe TPO-R. Accordingly, such peptides and peptide mimetics are usefulfor therapeutic purposes in treating conditions mediated by TPO (e.g.,thrombocytopenia resulting from chemotherapy, radiation therapy, or bonemarrow transfusions) as well as for diagnostic purposes in studying themechanism of hematopoiesis and for the in vitro expansion ofmegakarovcytes and committed progenitor cells.

Peptides and peptide mimetics suitable for therapeutic and/or diagnosticpurposes have an IC₅₀ of about 2 mM or less, as determined by thebinding affinity assay set forth in Example 3 below wherein a lower IC₅₀correlates to a stronger binding affinity to TPO-R. For pharmaceuticalpurposes, the peptides and peptidomimetics preferably have an IC₅₀ of nomore than about 100 μm, more preferably, no more than 500 nM. In apreferred embodiment, the molecular weight of the peptide or peptidemimetic is from about 250 to about 8000 daltons.

When used for diagnostic purposes, the peptides and peptide mimeticspreferably are labeled with a detectable label and, accordingly, thepeptides and peptide mimetics without such a label serve asintermediates in the preparation of labeled peptides and peptidemimetics.

Peptides meeting the defined criteria for molecular weight and bindingaffinity for TPO-R comprise 9 or more amino acids wherein the aminoacids are naturally occurring or synthetic (non-naturally occurring)amino acids. Peptide mimetics include peptides having one or more of thefollowing modifications:

peptides wherein one or more of the peptidyl [--C(O)NR--] linkages(bonds) have been replaced by a non-peptidyl linkage such as a --CH₂-carbamate linkage [--CH₂ --OC(O)NR--]; a phosphonate linkage; a --CH₂-sulfonamide [--CH₂ --S(O)₂ NR-] linkage; a urea [--NHC(O)NH--] linkage;a --CH₂ -secondary amine linkage; or an alkylated peptidyl linkage[--C(O)NR⁶ -- where R⁶ is lower alkyl];

peptides wherein the N-terminus is derivatized to a --NRR¹ group; to a--NRC(O)R group; to a --NRC(O)OR group; to a --NRS(O)₂ R group; to a--NHC(O)NHR group where R and R¹ are hydrogen or lower alkyl with theproviso that R and R¹ are not both hydrogen; to a succinimide group; toa benzyloxycarbonyl--NH-- (CBZ--NH--) group; or to abenzyloxycarbonyl--NH-- group having from 1 to 3 substituents on thephenyl ring selected from the grout consisting of lower alkyl, loweralkoxy, chloro, and bromo; or

peptides wherein the C terminus is derivatized to --C(O)R² where ² isselected from the group consisting of lower alkoxy, and --NR³ R⁴ whereR³ and R⁴ are independently selected from the group consisting ofhydrogen and lower alkyl.

Accordingly, preferred peptides and peptide mimetics comprise a compoundhaving:

(1) a molecular weight of less than about 5000 daltons, and

(2) a binding affinity to TPO-R as expressed by an IC₅₀ of no more thanabout 100μm,

wherein from zero to all of the --C(O)NH-- linkages of the peptide havebeen replaced by a linkage selected from the group consisting of a

--CH₂ OC(O)NR-- linkage; a phosphonate linkage; a --CH₂ S(O)₂ NR--linkage; a --CH₂ NR-- linkage; and a --C(O)NR⁶ -- linkage; and a--NHC(O)NH-- linkage where R is hydrogen or lower alkyl and R⁶ is loweralkyl,

further wherein the N-terminus of said peptide or peptide mimetic isselected from the group consisting of a --NRR¹ group; a --NRC(O)R group;a --NRC(O)OR group; a --NRS(O)₂ R group; --NHC(O)NHR group; asuccinimide group; a benzyloxycarbonyl--NH-- group; and abenzyloxycarbonyl--NH-- group having from 1 to 3 substituents on thephenyl ring selected from the group consisting of lower alkyl, loweralkoxy, chloro, and bromo, where R and R¹ are independently selectedfrom the group consisting of hydrogen and lower alkyl,

and still further wherein the C-terminus of said peptide or peptidemimetic has the formula --C(O)R² where R² is selected from the groupconsisting of hydroxy, lower alkoxy, and --NR³ R⁴ where R³ and R⁴ areindependently selected from the group consisting of hydrogen and loweralkyl and where the nitrogen atom of the --NR³ R⁴ group can optionallybe the amine group of the N-terminus of the peptide so as to form acyclic peptide,

and physiologically acceptable salts thereof.

In a related embodiment, the invention is directed to a labeled peptideor peptide mimetic comprising a peptide or peptide mimetic described asabove having covalently attached thereto a label capable of detection.

In some embodiments of the invention, preferred peptides for use includepeptides having a core structure comprising a sequence of amino acids(SEQ ID NO:2):

    X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7

where X₁ is C, L, M, P, Q, V; X₂ is F, K, L, N, Q, R, S, T or V; X₃ isC, F, I, L, M, R, S, V or W; X₄ is any of the 20 genetically codedL-amino acids; X₅ is A, D, E, G, K, M, Q, R, S, T, V or Y; X₆ is C, F,G, L, M, S, V, W or Y; and X₇ is C, G, I, K, L, M, N, R or V.

In a preferred embodiment the core peptide comrpises a sequence of aminoacids (SEQ ID NO:3):

    X.sub.8 G X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 W X.sub.7

where X₁ is L, M, P, Q. or V; X₂ is F, R, S, or T; X₃ is F, L, V, or W;X₄ is A, K, L, M, R, S, V, or T; X₅ is A, E, G, K, M, Q, R, S, or T; X₇is C, I, K, L, M or V; and each X₈ residue is independently selectedfrom any of the 20 genetically coded L-amino acids, their stereoisomericD-amino acids; and non-natural amino acids. Preferably, each X₈ residueis independently selected from any of the 20 genetically coded L-aminoacids and their stereoisomeric D-amino acids. In a preferred embodiment(SEQ ID NO:4), X₁ is P; X₂ is T; X₃ is L; X₄ is R; X₅ is E or Q; and X₇is I or L.

More preferably, the core peptide comprises a sequence of amino acids(SEQ ID NO:5):

    X.sub.9 X.sub.8 G X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 W X.sub.7

where X₉ is A, C, E, G, I, L , M, P, R, Q, S, T, or V; and X₈ is A, C,D, E, K, L, Q, R, S, T, or V. More preferably, X₉ is A or I; and X₈ isD, E, or K.

Particularly preferred peptides include (SEQ ID NOS:6-13, respectively):G G C A D G P T L R E W I S F C G G; G N A D G P T L R Q W L E G R R P KN; G G C A D G P T L R E W I S F C G G K; T I K G P T L R Q W L K S R EH T S; S I E G P T L R E W L T S R T P H S; L A I E G P T L R Q W L H GN G R D T; C A D G P T L R E W I S F C; and I E G P T L R Q W L A A R A.

In further embodiments of the invention, preferred peptides for use inthis invention include peptides having a core structure comprising asequence of amino acids (SEQ ID NO:14):

    C X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7

where X₂ is K, L, N, Q, R, S, T or V; X₃ is C, F, I, L, M, R, S or V; X₄is any of the 20 genetically coded L-amino acids; X₅ is A, D, E, G, S, Vor Y; X₆ is C, F, G, L, M, S, V, W or Y; and X₇ is C, G, I, K, L, M, N,R or V. In a more preferred embodiment, X₄ is A, E, G, H, K, L, M, P. Q,R, S, T, or W. In a further embodiment, X₂ is S or T; X₃ is L or R; X₄is R; X₅ is D, E, or G; X₆ is F, L, or W; and X₇ is I, K, L, R, or V.Particularly preferred peptides include (SEQ ID NO:15): G G C T L R E WL H G G F C G G.

In a further embodiment, preferred peptides for use in this inventioninclude peptides having a structure comprising a sequence of amino acids(SEQ ID NO:16):

    X.sub.8 C X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7

where X₂ is F, K, L, N, Q, R, S, T or V; X₃ is C, F, I, L, M, R, S, V orW; X₄ is any of the 20 genetically coded L-amino acids; X₅ is A, D, E,G, K, M, Q, R, S, T, V or Y; X₆ is C, F, G, L, M, S, V, W or Y; X₇ is C,G, I, K, L, M, N, R or V; and X₈ is any of the 20 genetically codedL-amino acids. In some embodiments, X₈ is preferably G, S, Y, or R.

The compounds described herein are useful for the prevention andtreatment of diseases mediated by TPO, and particularly for treatinghematological disorders, including but not limited to, thrombocytopeniaresulting from chemotherapy, radiation therapy, or bone marrowtransfusions. Thus, the present invention also provides a method fortreating wherein a patient having a disorder that is susceptible totreatment with a TPO agonist receives, or is administered, atherapeutically effective dose or amount of a compound of the presentinvention.

The invention also provides for pharmaceutical compositions comprisingone or more of the compounds described herein and a physiologicallyacceptable carrier. These pharmaceutical compositions can be in avariety of forms including oral dosage forms, as well as inhalablepowders and solutions and injectable and infusible solutions.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A-B illustrates the results of a functional assay in the presenceof various peptides; the assay is described in Example 2. FIG. 1A is agraphical depiction of the results of the TPO-R transfected Ba/F3 cellproliferation assay for selected peptides of the invention:

▪ designating the results for (SEQ ID NO:8) G G C A D G P T L R E W I SF C G G K (biotin);

X designating the results for (SEQ ID NO:6) G G C A D G P T L R E W I SF C G G;

▴ designating the results for (SEQ ID NO:11) L A I E G P T L R Q W L H GN G R D T;

◯ designating the results for G N A D G P T L R Q W L E G R R P K N; and

+ designating the results for (SEQ ID NO:9) T I K G P T L R Q W L K S RE H T S.

FIG. 1B is a graphical depiction of the results with the same peptidesand the parental cell line.

FIGS. 2A-C show the results of peptide oligomerization using the TPO-Rtransfected Ba/F3 cell proliferation assay. FIG. 2A shows the results ofthe assay for the complexed biotinylated peptide (AF 12285 withstreptavidin (SA)) for both the transfected and parental cell lines.FIG. 2B shows the results of the assay for the free biotinylated peptide(AF 12285) for both the transfected and parental cell lines. FIG. 2Cshows the results of the assay for streptavidin alone for both thetransfected and parental cell lines.

FIGS. 3A-G show the results of a series of control experiments showingthe activity of TPO, the peptides of the present invention, EPO, andEPO-R binding peptides in a cell proliferation assay using either theTPO-R transfected sa/F3 cell line and its corresponding parental line,or an EPO-dependent cell line. FIG. 3A depicts the results for TPO inthe cell proliferation assay using the TPO-R transfected Ba/F3 cell lineand its corresponding parental line. FIG. 3B depicts the results for EPOin the cell proliferation assay using the TPO-R transfected Ba/F3 cellline and its corresponding parental line. FIG. 3C depicts the resultsfor complexed biotinylated peptide (AF 12285 with streptavidin (SA)) anda complexed form of a biotinylated EPO-R binding peptide (AF 11505 withSA) in the TPO-R transfected Ba/F3 cell line. The results for thecorresponding parental cell line are shown in FIG. 3D. FIG. 3E depictsthe results for TPO in the cell, proliferation assay using theEPO-dependent cell line. FIG. 3F depicts the results for EPO in the cellproliferation assay using the EPO-dependent cell line. FIG. 3G depictsthe results for complexed biotinylated peptide (AF 12885 withstreptavidin (SA)) and the complexed form of a biotinylated EPO-Rbinding peptide (AF 11505 with SA) in the EPO-dependent cell line.

FIGS. 4A-C illustrates the construction of peptides-on-plasmidslibraries in vector pJS142. FIG. 4A shows a restriction map and positionof the genes. The library plasmid includes the rrnB transcriptionalterminator, the bla gene to permit selection on ampicillin, the M13phage intragenic region (M13 IG) to permit rescue of single-strandedDNA, a plasmid replication origin (ori) , two lacO_(s) sequences, andthe araC gene to permit positive and negative regulation of the araBpromoter driving expression of the lac fusion gene. FIG. 4B (SEQ IDNOS:19 -20, respectively) shows the sequence of the cloning region atthe 3' end of the lac I gene, including the SfiI and EagI sites usedduring library construction. FIG. 4C (SEQ ID NOS:223-224, respectively)shows the ligation of annealed library oligonucleotides, ON-829 andON-830, to SfiI sites of pJS142 to produce a library. Single spaces inthe sequence indicate sites of ligation.

FIGS. 5A-B illustrate cloning into the pELM3 and pELM15 MBP vectors.FIG. 5A (SEQ ID NOS:225-226, respectively) shows the sequence at the 3'end of the malE fusion gene, including the MBP coding sequence, the polyasuaraaine linker, the factor Xa protease cleavagge site, and theavailable cloning sites. The remaining portions of the vectors arederived from pMALc2 (pELM3) and pMALp2 (pELM15), available from NewEngland Biolabs. FIG. 5B (SEQ ID NOS:227-228, respectively) shows thesequence of the vectors after transfer of the BspEII-ScaI libraryfragment into AgeI-ScaI digested pELM3/pELM15. The transferred sequenceincludes the sequence encoding the GGG peptide linker from the pJS142library.

FIG. 6A depicts a restriction map and position of the genes for theconstruction of headpiece dimer libraries in vector pCMG14. The libraryplasmid includes: the rrnB transcriptional terminator, the bla gene topermit selection on ampicillin, the M13 phage intragenic region (M13 IG)to permit rescue of single-scranded DNA, a plasmid replication origin(ori), one lacO_(s) ssequence, and the araC gene to permit positive andnegative regulation of the araB promoter driving expression of theheadpiece dimer fusion gene. FIG. 6B (SEQ ID NOS:229-230, respectively)depicts the sequence of the cloning region at the 3' end of theheadpiece dimer gene, including the SfiI and EagI sites used duringlibrary construction. FIG. 6C (SEQ ID NOS:231-232, respectively) showsthe ligation of annealed ON-1679, ON-829, and ON-830 to SfiI sites ofpCMG14 to produce a library. Singles spaces in the sequence indicatesites of ligation.

FIGS. 7 to 9 show the results of further assays evaluating activity ofthe peptides and peptide mimETICS of the invention. In this assay miceare made thrombocytopenic with carboplatin. FIG. 7 depicts typicalresults when Balb/C mice are treated with carboplatin (125 mg/kgintraperitoneally) on Day 0. The dashed lines represent untreatedanimals from three experiments. The solid line representcarboplatin-treated groups in three experiments. The heavy solid linesrepresent historical data. FIG. 8 depicts the effect of carboplatintitration on platelet counts in mice treated with the indicated amountsof carboplatin (in mg/kg, intraperitoneally (ip) on Day 0). FIG. 9depicts amelioration of carboplatin-induced thrombocytopenia on Day 10by peptide AF12513 (513). Carboplatin (CBP; 50-125 mg/kg,intraperitoneally) was administered on Day 0. AF12513 (1 mg/kg, ip) wasgiven on Days 1-9.

DESCRIPTION OF SPECIFIC EMBODIMENTS

I. DEFINITIONS AND GENERAL PARAMETERS

The following definitions are set forth to illustrate and define themeaning and scope of the various terms used to describe the inventionherein.

"Agonist" refers to a biologically active ligand which binds to itscomplementary biologically active receptor and activates the lattereither to cause a biological response in the receptor or to enhancepreexisting biological activity of the receptor.

"Pharmaceutically acceptable salts" refer to the non-toxic alkali metal,alkaline earth metal, and ammonium salts commonly used in thepharmaceutical industry including the sodium, potassium, lithium,calcium, magnesium, barium, ammonium, and protamine zinc salts, whichare prepared by methods well known in the art. The term also includesnon-toxic acid addition salts, which are generally prepared by reactingthe compounds of this invention with a suitable organic or inorganicacid. Representative salts include the hydrochloride, hydrobromide,sulfate, bisulfate, acetate, oxalate, valerate, oleate, laurate, borate,benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate,succinate, tartrate, napsylate, and the like.

"Pharmaceutically acceptable acid addition salt" refers to those saltswhich retain the biological effectiveness and properties of the freebases and which are not biologically or otherwise undesirable, formedwith inorganic acids such as hydrochloric acid, hydrobromic acid,sulfuric acid, nitric acid, phosphoric acid and the like, and organicacids such as acetic acid, propionic acid, glycolic acid, pyruvic acid,oxalic acid, malic acid, malonic acid, succinic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid,mandelic acid, menthanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid and the like. For a descriptionof pharmaceutically acceptable acid addition salts as prodrugs, seeBundgaard, H., supra.

"Pharmaceutically acceptable ester" refers to those esters which retain,upon hydrolysis of the ester bond, the biological effectiveness andproperties of the carboxylic acid or alcohol and are not biologically orotherwise undesirable. For a description of pharmaceutically acceptableesters as prodrugs, see Bundgaard, H., ed., Design of Prodrugs, ElsevierScience Publishers, Amsterdam (1985). These esters are typically formedfrom the corresponding carboxylic acid and an alcohol. Generally, esterformation can be accomplished via conventional synthetic techniques.(See, e.g., March Advanced Organic Chemistry, 3rd Ed., John Wiley &Sons, New York (1985) p. 1157 and references cited therein, and Mark etal. Encyclopedia of Chemical Technology, John Wiley & Sons, New York(1980)). The alcohol component of the ester will generally comprise (i)a C₂ -C₁₂ aliphatic alcohol that can or can not contain one or moredouble bonds and can or can not contain branched carbons or (ii) a C₇-C₁₂ aromatic or heteroaromatic alcohols. This invention alsocontemplates the use of those compositions which are both esters asdescribed herein and at the same time are the pharmaceuticallyacceptable acid addition salts thereof.

"Pharmaceutically acceptable amide" refers to those amides which retain,upon hydrolysis of the amide bond, the biological effectiveness andproperties of the carboxylic acid or amine and are not biologically orotherwise undesirable. For a description of pharmaceutically acceptableamides as prodrugs, see Bundgaard, H., ed., Design of Prodrugs, ElsevierScience Publishers, Amsterdam (1985). These amides are typically formedfrom the corresponding carboxylic acid and an amine. Generally, amideformation can be accomplished via conventional synthetic techniques.(See, e.g., March Advanced Organic Chemistry, 3rd Ed., John Wiley &Sons, New York (1985) p. 1152 and Mark et al. Encyclopedia of ChemicalTechnology, John Wiley & Sons, New York (1980)) This invention alsocontemplates the use of those compositions which are both amides asdescribed herein and at the same time are the pharmaceuticallyacceptable acid addition salts thereof.

"Pharmaceutically or therapeutically acceptable carrier" refers to acarrier medium which does not interfere with the effectiveness of thebiological activity of the active ingredients and which is not toxic tothe host or patient.

"Stereoisomer" refers to a chemical compound having the same molecularweight, chemical composition, and constitution as another, but with theatoms grouped differently,. That is,. certain identical chemicalmoieties are at different orientations in space and, therefore, whenpure, has the ability to rotate the plane of polarized light. However,some pure stereoisomers may have an optical rotation that is so slightthat it is undetectable with present instrumentation. The compounds ofthe instant invention may have one or more asymmetrical carbon atoms andtherefore include various stereoisomers. All stereoisomers are includedwithin the scope of the invention.

"Therapeutically- or pharmaceutically-effective amount" as applied tothe compositions of the instant invention refers to the amount ofcomposition sufficient to induce a desired biological result. Thatresult can be alleviation of the signs, symptoms, or causes of adisease, or any other desired alteration of a biological system. In thepresent invention, the result will typically involve a decrease in theimmunological and/or inflammatory responses to infection or tissueinjury.

Amino acid residues in peptides are abbreviated as follows:Phenylalanine is Phe or F; Leucine is Leu or L; Isoleucine is Ile or I;Methionine is Met or M; Valine is Val or V; Serine is Ser or S; Prolineis Pro or P; Threonine is Thr or T; Alanine is Ala or A; Tyrosine is Tyror Y; Histidine is His or H; Glutamine is Gln or Q; Asparagine is Asn orN; Lysine is Lys or K; Asparcic Acid is Asp or D; Glutamic Acid is Gluor E; Cysteine is Cys or C; Tryptophan is Trp or W; Arginine is Arg orR; and Glycine is Gly or G. Additionally, Bu is Butoxy, Bzl is benzyl,CHA is cyclohexylamine, Ac is acetyl, Me is methyl, Pen ispenicillamine, Aib is amino isobutyric acid, Nva is norvaline, Abu isamino butyric acid, Thi is thienylalanine, OBn is O-benzyl, and hyp ishydroxyproline.

In addition to peptides consisting only of naturally-occurring aminoacids, peptidomimetics or peptide analogs are also provided. Peptideanalogs are commonly used in the pharmaceutical industry as non-peptidedrugs with properties analogous to those of the template peptide. Thesetypes of non-peptide compound are termed "peptide mimetics" or"peptidomimetics" (Fauchere, J. Adv. Drug Res. 15:29 (1986); Veber andFreidinger TINS p.392 (1985); and Evans et al. J. Med. Chem. 30:1229(1987), which are incorporated herein by reference). Peptide mimeticsthat are structurally similar to therapeutically useful peptides may beused to produce an equivalent or enhanced therapeutic or prophylacticeffect. Generally, peptidomimetics are structurally similar to aparadigm polypeptide (i.e., a polypeptide that has a biological orpharmacological activity), such as naturally-occurring receptor-bindingpolypeptide, but have one or more peptide linkages optionally replacedby a linkage selected from the group consisting of: --CH₂ NH--, --CH₂S--, --CH₂ --CH₂ --, --CH=CH-- (cis and trans), --COCH₂ --, --CH(OH)CH₂--, and --CH₂ SO--, by methods known in the art and further described inthe following references: Spatola, A. F. in Chemistry and Biochemistryof Amino Acids, Peptides, and Proteins, B. Weinstein, eds., MarcelDekker, New York, p. 267 (1983); Spatola, A. F., Vega Data (March 1983),Vol. 1, Issue 3, Peptide Backbone Modifications (general review);Morley, Trends Pharm Sci (1980) pp. 463-468 (general review); Hudson, D.et al., Int J Pept Prot Res 14:177-185 (1979) (--CH₂ NH--, CH₂ CH₂ --);Spatola et al. Life Sci 38:1243-1249 (1986) (--CH₂ --S); Hann J. Chem.Soc Perkin Trans. I 307-314 (1982) (--CH--CH--, cis and trans); Almquistet al. J. Med. Chem. 23:1392-1398 (1980) (--COCH₂ --); Jennings-White etal. Tetrahedron Lett 23:2533 (1982) (--COCH₂ --); Szelke et al. EuropeanAppln. EP 45665 CA (1982): 97:39405 (1982) (--CH(OH)CH₂ --); Holladay etal. Tetrahedron Lett 24:4401-4404 (1983) (--C(OH)CH₂ --); and Hruby LifeSci 31:189-199 (1982) (--CH₂ --S--); each of which is incorporatedherein by reference. A particularly preferred non-peptide linkage is--CH₂ NH--. Such peptide mimetics may have significant advantages overpolypeptide embodiments, including, for example: more economicalproduction, greater chemical stability, enhanced pharmacologicalproperties (half-life, absorption, potency, efficacy, etc.), alteredspecificity (e.g., a broad-spectrum of biological activities), reducedantigenicity, and others. Labeling of peptidomimetics usually involvescovalent attachment of one or more labels, directly or through a spacer(e.g., an amide group); to non-interfering position(s) on thepeptidomimetic that are predicted by quantitative structure-activitydata and/or molecular modeling. Such non-interfering positions generallyare positions that do not form direct contacts with themacromolecules(s) (e.g., immunoglobulin superfamily molecules) to whichthe peptidomimetic binds to produce the therapeutic effect.Derivitization (e.g., labeling) of peptidomimetics should notsubstantially interfere with the desired biological or pharmacologicalactivity of the peptidomimetic. Generally, peptidomimetics ofreceptor-binding peptides bind to the receptor with high affinity andpossess detectable biological activity (i.e., are agonistic orantagonistic to one or more receptor-mediated phenotypic changes).

Systematic substitution of one or more amino acids of a consensussequence with a D-amino acid of the same type (e.g., D-lysine in placeof L-lysine) may be used to generate more stable peptides. In addition,constrained peptides comprising a consensus sequence or a substantiallyidentical consensus sequence variation may be generated by methods knownin the art (Rizo and Gierasch Ann. Rev. Biochem. 61:387 (1992),incorporated herein by reference); for example, by adding internalcysteine residues capable of forming intramolecular disulfide bridgeswhich cyclize the peptide.

Synthetic or non-naturally occuring amino acids refer to amino acidswhich do not naturally occur in vivo but which, nevertheless, can beincorporated into the peptide structures described herein. Preferredsynthetic amino acids are the D-a-amino acids of naturally occurringL-a-amino acid as well as non-naturally occurring D- and L-a-amino acidsrepresented by the formula H₂ NCHR⁵ COOH where R⁵ is 1) a lower alkylgroup, 2) a cycloalkyl group of from 3 to 7 carbon atoms, 3) aheterocycle of from 3 to 7 carbon atoms and 1 to 2 heteroatoms selectedfrom the group consisting of oxygen, sulfur, and nitrogen, 4) anaromatic residue of from 6 to 10 carbon atoms optionally having from 1to 3 substituents on the aromatic nucleus selected from the groupconsisting of hydroxyl, lower alkoxy, amino, and carboxyl, 5)-alkylene-Y where alkylene is an alkylene group of from 1 to 7 carbonatoms and Y is selected from the group consisting of (a) hydroxy, (b)amino, (c) cycloalkyl and cycloalkenyl of from 3 to 7 carbon atoms, (d)aryl of from 6 to 10 carbon atoms optionally having from 1 to 3substituents on the aromatic nucleus selected from the group consistingof hydroxyl, lower alkoxy, amino and carboxyl, (e) heterocyclic of from3 to 7 carbon atoms and 1 to 2 heteroatoms selected from the groupconsisting of oxygen, sulfur, and nitrogen, (f) --C(O)R² where R² isselected from the group consisting of hydrogen, hydroxy, lower alkyl,lower alkoxy, and --NR³ R⁴ where R³ and R⁴ are independently selectedfrom the group consisting of hydrogen and lower alkyl, (g) --S(O)_(n) R⁶where n is an integer from 1 to 2 and R⁶ is lower alkyl and with theproviso that R⁵ does not define a side chain of a naturally occurringamino acid.

Other preferred synthetic amino acids include amino acids wherein theamino group is separated from the carboxyl group by more than one carbonatom such as b-alanine, g-aminobutyric acid, and the like.

Particularly preferred synthetic amino acids include, by way of example,the. D-amino acids of naturally occurring L-amino acids,L-1-napthyl-alanine, L-2-naphthylalanine, L-cyclohexylalanine, L-2-aminoisobutyric acid, the sulfoxide and sulfone derivatives of methionine(i.e., HOOC-(H₂ NCH)CH₂ CH₂ -S(O)_(n) R⁶) where n and R₆ are as definedabove as well as the lower alkoxy derivative of methionine (i.e.,HOOC-(H₂ NCH)CH₂ CH₂ --OR⁶ where R⁶ is as defined above).

"Detectable label" refers to materials, which when covalently attachedto the peptides and peptide mimetics of this invention, permit detectionof the peptide and peptide mimetics in vivo in the patient to whom thepeptide or peptide mimetic has been administered. Suitable detectablelabels are well known in the art and include, by way of example,radioisotopes, fluorescent labels (e.g., fluorescein), and the like. TheDarticular detectable label employed is not critical and is selectedrelative to the amount of label to be employed as well as the toxicityof the label at the amount of label employed. Selection of the labelrelative to such factors is well within the skill of the art.

Covalent attachment of the detectable label to the peptide or peptidemimetic is accomplished by conventional methods well known in the art.For example, when the ¹²⁵ I radioisotope is employed as the detectablelabel, covalent attachment of ¹²⁵ I to the peptide or the peptidemimetic can be achieved by incorporating the amino acid tyrosine intothe peptide or peptide mimetic and then iodating the peptide. Iftyrosine is not present in the peptide or peptide mimetic, incorporationof tyrosine to the N or C terminus of the peptide or peptide mimetic canbe achieved by well known chemistry. Likewise, ³² P can be incorporatedonto the peptide or peptide mimetic as a phosphate moiety through, forexample, a hydroxyl group on the peptide or peptide mimetic usingconventional chemistry.

II. OVERVIEW

The present invention provides compounds that bind to and activate theTPO-R or otherwise behave as a TPO agonist. These compounds include"lead" peptide compounds and "derivative" compounds constructed so as tohave the same or similar molecular structure or shape as the leadcompounds but that differ from the lead compounds either with respect tosusceptibility to hydrolysis or proteolysis and/or with respect to otherbiological properties, such as increased affinity for the receptor. Thepresent invention also provides compositions comprising an effectiveamount of a TPO agonist, and more particularly a compound, that isuseful for treating hematological disorders, and particularly,thrombocytopenia associated with chemotherapy, radiation therapy, orbone marrow transfusions.

III. IDENTIFICATION OF TPO-AGONISTS

Peptides having a binding affinity to TPO-R can be readily identified byrandom peptide diversity generating systems coupled with an affinityenrichment process.

Specifically, random peptide diversity generating systems include the"peptides on plasmids" system described in U.S. Pat. Nos. 5,270,170 and5,338,665; the "peptides on phage" system described in U.S. patentapplication Ser. No. 07/718,577, filed Jun. 20, 1991 which is acontinuation in part application of U.S. patent application Ser. No.37/541,108, filed Jun. 20, 1990, and in Cwirla et al., Proc. Natl. Acad.Sci. USA 87:6378-6382 (1980); the "polysome system" described in U.S.patent application Ser. No. 08/300,262, filed Sep. 2, 1994, which is acontinuation-in-part application based on U.S. patent application Ser.No. 08/144,775, filed Oct. 29, 1993 and PCT WO 95/11992; the "encodedsynthetic library (ESL)" system described in U.S. patent applicationSer. No. 08/146,886, filed Nov. 12, 1993 which is a continuation in partapplication of U.S. patent application Ser. No. 07/946,239, filed Sep.16, 1992, which is a continuation in part application of U.S. patentapplication Ser. No. 07/762,522, filed Sep. 18, 1991; and the "verylarge scale immobilized polymer synthesis" system described in U.S. Pat.No. 5,143,854; PCT Patent Publication No. 90/15070, published Dec. 13,1990; U.S. patent application Ser. No. 07/624,120, filed Dec. 6, 1990;Fodor et al. Science 251:767-773 (2/1991); Dower and Fodor Ann. Rep.Med. Chem. 26:271-180 (1991); and U.S. patent application Ser. No.805,727, filed Dec. 6, 1991.

Using the procedures described above, random peptides were generallydesigned to have a defined number of amino acid residues in length(e.g., 12). To generate the collection of oligonucleotides encoding therandom peptides, the codon motif (NNK)x, where N is nucleotide A, C, G,or T (equimolar; depending on the methodology employed, othernucleotides can be employed), K is G or T (equimolar), and x is aninteger corresponding to the number of amino acids in the peptide (e.g.,12) was used to specify any one of the 32 possible codons resulting fromthe NNK motif: 1 for each of 12 amino acids, 2 for each of 5 aminoacids, 3 for each of 3 amino acids, and only one of the three stopcodons. Thus, the NNK motif encodes all of the amino acids, encodes onlyone stop codon, and reduces codon bias.

In the systems employed, the random peptides were presented either onthe surface of a phage particle, as part of a fusion protein comprisingeither the pIII or the pVIII coat protein of a phage fd derivative(peptides on phage) or as a fusion protein with the LacI peptide fusionprotein bound to a plasmid (peptides on plasmids).

The phage or plasmids, including the DNA encoding the peptides, wereidentified and isolated by an affinity enrichment process usingimmobilized TPO-R. The affinity enrichment process, sometimes called"panning," involves multiple rounds of incubating the phage, plasmids,or polysomes with the immobilized receptor, collecting the phage,plasmids, or polysomes that bind to the receptor (along with theaccompanying DNA or mRNA), and producing more of the phage or plasmids(along with the accompanying LacI-peptide fusion protein) collected. Theextracellular domain (ECD) of the TPO-R typically was used duringpanning.

After several rounds of affinity enrichment, the phage or plasmids andaccompanying peptides were examined by ELISA to determine if thepeptides bind specifically to TPO-R. This assay was carried outsimilarly to the procedures used in the affinity enrichment process,except that after removing unbound phage, the wells were typicallytreated with rabbit anti-phage antibody, then with alkaline phosphatase(AP)-conjugated goat anti-rabbit antibody. The amount of alkalinephosphatase in each well was determined by standard methods. A similarELISA procedure for use in the peptides on plasmids system is describedin detail below.

By comparing test wells with control wells (no receptor), one candetermine whether the fusion proteins bind to the receptor specifically.The phage pools found to bind to TPO-R were screened in a colony liftprobing format using radiolabelled monovalent receptor. This probe canbe produced using protein kinase A to phosphorylate a kemptide sequencefused to the C-terminus of the soluble receptor. The "engineered" formof the TPO receptor is then expressed in host cells, typically CHOcells. Following PI-PLC harvest of the receptors, the receptor wastested for binding to TPO or TPO-R specific phage clones. The receptoris then labeled to high specific activity with ³³ P for use as amonovalent probe to identify high affinity ligands using colony lifts.

Peptides found to bind specifically to the receptor were thensynthesized as the free peptide (e.g., no phage) and tested in ablocking assay. The blocking assay was carried out in similar fashion tothe ELISA , except that TPO or a reference peptide was added to thewells before the fusion protein (the control wells were of two types:(1) no receptor; and (2) no TPO or reference peptide). Fusion proteinsfor which the binding to the receptor was blocked by TPO or thereference peptide contain peptides in the random peptide portion thatare preferred compounds of the invention.

TPO-R, as well as its extracellular domain, were produced in recombinanthost cells. One useful form of TPO-R is constructed by expressing theprotein as a soluble protein in baculovirus transformed host cells usingstandard methods; another useful form is constructed with a signalpeptide for protein secretion and for glycophospholipid membrane anchorattachment. This form of anchor attachment is called "PIG-tailing". SeeCaras and Wendell Science 243:1196-1198 (1989) and Lin et al. Science249:677-679 (1990).

Using the PIG-tailing system, one can cleave the receptor from thesurface of the cells expressing the receptor (e.g., transformed CHOcells selected for high level expression of receptor with a cell sorter)with phospholipase C. The cleaved receptor still comprises a carboxyterminal sequence of amino acids, called the "HPAP tail", from thesignal protein for membrane attachment and can be immobilized withoutfurther purification. The recombinant receptor protein can beimmobilized by coating the wells of microtiter plates with an anti-HPAPtail antibody (Ab 179 or MAb 179), blocking non-specific binding withbovine serum albumin (BSA) in PBS, and then binding cleaved recombinantreceptor to the antibody. Using this procedure, one should perform theimmobilization reaction in varying concentrations of receptor todetermine the optimum amount for a given preparation, because differentpreparations of recombinant protein often contain different amounts ofthe desired protein. In addition, one should ensure that theimmobilizing antibody is completely blocked (with TPO or some otherblocking compound) during the affinity enrichment process. Otherwise,unblocked antibody can bind undesired phage during the affinityenrichment procedure. One can use peptides that bind to the immobilizingantibody to block unbound sites that remain after receptorimmobilization to avoid this problem or one can simply immobilize thereceptor directly to the wells of microtiter plates, without the aid ofan immobilizing antibody. See U.S. patent application Ser. No.07/947,339, filed Sep. 18, 1992, incorporated herein by reference.

When using random peptide generation systems that allow for multivalentligand-receptor interaction, one must recognize that the density of theimmobilized receptor is an important factor in determining the affinityof the ligands that can bind to the immobilized receptor. At higherreceptor densities (e.g., each anti-receptor antibody-coated welltreated with 0.25 to 0.5 mg of receptor), multivalent binding is morelikely to occur than at lower receptor densities (e.g., eachanti-receptor antibody-coated well treated with 0.5 to 1 ng of thereceptor). If multivalent binding is occurring, then one will be morelikely to isolate ligands with relatively lower affinity, unless oneuses high densities of immobilized receptor to identify lead compoundsand uses lower receptor densities to isolate higher affinity derivativecompounds.

To discriminate among higher affinity peptides, a monovalent receptorprobe frequently is used. This probe can be produced using proteinkinase A to phosphorylate a kemptide sequence fused to the C-terminus ofthe soluble receptor. The "engineered" form of the TPO receptor is thenexpressed in host cells, typically CHO cells. Following PI-PLC harvestof the receptors, the receptor was tested for binding to TPO or TPO-Rspecific phage clones. The receptor is then labeled to high specificactivity with ³³ P for use as a monovalent probe to identify highaffinity ligands using colony lifts.

Preferred screening methods to facilitate identification of peptideswhich bind TPO-R involve first identifying lead peptides which bind tothe extracellular domain of the receptor and then making other peptideswhich resemble the lead peptides. Specifically, using a pIII orpVIII-based peptides on phage system, a random library can be screenedto discover a phage that presents a peptide that binds to TPO-R. Thephage DNAs are sequenced to determine the sequences of the peptidesdisplayed on the surface of the phages.

Clones capable of specific binding to the TPO-R were identified from arandom linear 10-mer pVIII library and a random cyclic 10-mer and 12-merpVIII libraries. The sequences of these peptides serve as the basis forthe construction of other peptide libraries designed to contain a highfrequency of derivatives of the initially identified peptides. Theselibraries can be synthesized so as to favor the production of peptidesthat differ from the binding peptide in only a few residues. Thisapproach involves the synthesis of an oligonucleotide with the bindingpeptide coding sequence, except that rather than using pure preparationsof each of the four nucleotide triphosphates in the synthesis, one usesmixtures of the four nucleoside triphosphates (i.e., 55% of the"correct" nucleotide, and 15% each of the other three nucleotides is onepreferred mixture for this purpose and 70% of the "correct" nucleotideand 10% of each of the other three nucleotides is another preferredmixture for this purpose) so as to generate derivatives of the bindingpeptide coding sequence.

A variety of strategies were used to derivatize the lead peptides bymaking "mutagenesis on a theme" libraries,. These included a pVIIIphagemid mutagenesis library based on the consensus sequence mutagenizedat 70:10:10:10 frequency and extended on each terminus with randomresidues to produce clones which enclode the sequence (SEQ ID NO:21)XXXX (C, S, P, or R) TLREWL XXXXXX (C or S). A similarextended/mutagenized library was constructed using thepeptides-on-plasmids system to produce clones which enclode the sequence(SEQ ID NO:21) XXXXX (C, S, P, or R) TLREWL XXXXXXX. An additionalextended/mutagenized library (SEQ ID NO:23), XXXX (C, S, P, or R) TLREWLXXXXXX (C or S), was constructed using the polysome display system. Allthree libraries were screened with peptide elution and probed withradiolabeled monovalent receptor.

The "peptides on plasmids" techniques was also used for peptidescreening and mutagenesis studies and is described in greater detail inU.S. Pat. No. 5,338,665, which is incorporated herein by reference forall purposes. According to this approach, random peptides are fused atthe C-terminus of LacI through expression from a plasmid vector carryingthe fusion gene. Linkage of the LacI-peptide fusion to its encoding DNAoccurs via the lacO sequences on the plasmid, forming a stablepeptide-LacI-plasmid complex that can be screened by affinitypurification (panning) on an immobilized receptor. The plasmids thusisolated can then be reintroduced into E. coli by electroporation toamplify the selected population for additional rounds of screening, orfor the examination of individual clones.

In addition, random peptide screening and mutagenesis studies wereperformed using a modified C-terminal Lac-I display system in whichdisplay valency was reduced ("headpiece dimer" display system). Thelibraries were screened and the resulting DNA inserts were cloned as apool into a maltose binding protein (MBP) vector allowing theirexpression as a C-terminal fusion protein. Crude cell lysates fromrandomly picked individual MBP fusion clones were then assayed for TPO-Rbinding in an ELISA format, as discussed above.

Peptide mutagenesis studies were also conducted using the polysomedisplay system, as described in co-pending application U.S. patentapplication Ser. No. 08/300,262, filed Sep. 2, 1994, which is acontinuation-in-part application based on U.S. patent application Ser.No. 08/144,775, filed Oct. 29, 1993 and PCT WO 95/11992, each of whichis incorporated herein by references for all purposes. A mutagenesislibrary was constructed based on the sequence (SEQ ID NO:24) X X X X(C,P,R, or S) t l r e f l X X X X X X (C or S), in which X represents arandom NNK codon, and the lower case letters represent amino acid codonscontaining 70:10:10:10 mutagenesis at positions 1 and 2 and K (G or T)at position 3 of the codon. The library was panned for 5 rounds againstTPO receptor which had been immobilzed on magnetic beads. After thefifth round, the PCR amplified pool was cloned into pAFF6 and the ELISApositive clones were sequenced. The sequences were subcloned into an MBPvector and their binding affinities were determined by an MBP ELISA.

To imobilize the TPO-R for polysome screening, Ab 179 was firstchemically conjugated to tosyl-activated magnetic beads (available fromDynal Corporation) as described by the manufacturer. The beads wereincubated with antibody in 0.5 M borate buffer (pH 9.5) overnight atroom temperature. The beads were washed and combined with TPO-Rcontaining the "HPAP" tail. The antibody coated beads and receptor wereincubated for 1 hour at 4° C., and the beads were washed again prior toadding the polysome library.

Screening of the various libraries described above yielded the TPOreceptor binding peptides shown in Tables 1 and 2 below, as well asothers not listed herein.

                  TABLE 1                                                         ______________________________________                                        (SEQ ID NOS 25-58, respectively)                                              Peptide                                                                       ______________________________________                                        R E G P T L R Q W M                                                           R E G P T L R Q W M                                                           S R G M T L R E W L                                                           E G P T L R G W L A                                                           R E G Q T L K E W L                                                           E R G P F W A K A C                                                           R E G P R C V M W M                                                           C S G L T L R E W L V C                                                       C L T G P F V T Q W L Y E C                                                   C G E G L T L T Q W L E H C                                                   C R A G P T L L E W L T L C                                                   C R A G P T L L E W L T L C                                                   C R Q G P T L T A W L L E C                                                   C A D G P T L R E W I S F C                                                   C E L V G P S L M S W L T C                                                   C G T E G P T L S T W L D C                                                   C D Q L G V T L S R W L E C                                                   S G T G L T L R E W L G S F S L L S                                           C P E G P T L L Q W L K R G Y S S C                                           R G D G P T L S Q W L Y S L M I M C                                           M V A G P T L R E F I A S L P I H C                                           S M Q G P T F R E W V S M M K V L C                                           S V Q C G P T L R Q W L A A R N H L S                                         G N A D G P T L R Q W L E G R R P K N                                         S V R C G P T L R Q W L A A R T H L S                                         L A I E G P T L R Q W L H G N G R D T                                         H G R V G P T L R E W K T Q V A T K K                                         C A D G P T L R E W I S F C                                                   I S D G P T L K E W L S V T R G A S                                           S I E G P T L R E W L T S R T P H S                                           T I K G P T L R Q W L K S R E H T S                                           G N A D G P T L R Q W L E G R R P K N                                         S I E G P T L R E W L T S R T P H S                                           I S D G P T L K E W L S V T R G A S                                           ______________________________________                                    

                  TABLE 2                                                         ______________________________________                                        (SEQ ID NOS 59-167, respectively                                              Peptide                                                                       ______________________________________                                        C S L E D L R K R C                                                           C R R S E L L E R C                                                           C T F K Q F L D G C                                                           C T R G E W L R C C                                                           C T L R Q W L Q G C                                                           C T L E E L R A C C                                                           C T R E E L M R L C                                                           C Q R A D L I N F C                                                           C N R N D L L L F C                                                           C T R T E W L H G C                                                           C S L G E L R R L C                                                           C N I N Q L R S I C                                                           C T M R Q F L V C C                                                           C T R S E W L E R C                                                           C T L H E Y L S G C                                                           C T R E E L L R Q C                                                           C T F R E F V N G C                                                           C S R A D F L A A C                                                           C S C A Q V V Q C C                                                           C T L R Q W I L L G M C                                                       C T L R E W L H G G F C                                                       C T L R A W L M S E T C                                                       C T L R A W L M E S C C                                                       C T F Q V W K L A R N C                                                       C L L R E W L D X R T C                                                       C V L R E W L L X X S C                                                       C L L S E F L A G Q Q C                                                       C S L R Q Y L D F G L G S C                                                   C T L Q E L K Q S S L Y E C                                                   C D L S E L K T H G Y A Y C                                                   C K L S D W L M N G V A A C                                                   C S L Q E F L S H G G Y V C                                                   C S L K E F L H S G L M Q C                                                   C T F R Q L L E Y G V S S C                                                   C T M R E F L V A S G V A C                                                   C T L A E F L A S G V E Q C                                                   C T L A E F L A S G V E Q C                                                   C T L K E W L V S H E V W C                                                   C T L R E F L S L G M N A C                                                   C T L R E F L D P T T A V C                                                   C S L L E F L A L G V A L C                                                   G G G R G C T L K Q W K Q G D C G R S                                         C N R S Q L L A A C                                                           C T L Q Q W L S G C                                                           C T L R E F K A G C                                                           C T R A Q F L K G C                                                           C T L R E F N R G C                                                           C T L S D F K R G C                                                           C T F R Q W K E A C                                                           C T L S E F R G G C                                                           C T L Q E F L E G C                                                           C T L Q Q W K D G C                                                           C T R S Q W L E G C                                                           C S L Q E F K H G C                                                           C T L G E W K R G C                                                           C T L W G C G K R G C                                                         C T L Q E W R G G C                                                           C T R L S G C W L C                                                           C T R T Q W L L D C                                                           C T L A E F R R G C                                                           C T S T Q W L L A C                                                           C S R S Q F L R S C                                                           C T L R E W L E G C                                                           C T L R E F L L M G A C                                                       C T L K E W L L W S S C                                                       C T L L E W L R N P V C                                                       C T L R Q W L G D A W C                                                       C T L G Q W L Q M G M C                                                       C T L R E W V F A G L C                                                       C L L L E F L S G A D C                                                       C T L G E F L A G H L C                                                       C R L R E F L V D L T C                                                       C S F R S W L V D Q T C                                                       C T L R E W L E D L G C                                                       C T L Q D W L V S W T C                                                       C T L S E W L S E L S C                                                       C T L M Q W L G G W P C                                                       C T L R E W L S Y G T C                                                       C T L Q E W L S G G L C                                                       G S H G C T L R E W L C M K I V P C                                           Q W Q G C T L R D C I L R G V F W S                                           S V N S C T L R E F L T G C R V F C                                           S Y D G C T L R H W L M D I Y G D C                                           Q R S G C T L R D W V L L N C L A S                                           N Y R G C T L S Q W V S E Q I V G C                                           G R S G C T L R E Y L G G M C Y L S                                           A S W Y C T V P E L M E M Q L P E C                                           G S T G C T L R E X L H M L G L D C                                           A C E G C T L R Q W L E Y V R V G C                                           A Q R G C T L Q Y F V S Y G X D M C                                           G V C G C T L R E F L A I P H T S C                                           S E G G C T L R E W V A S S L A N C                                           S N S R C T L R E W I I Q G C D F S                                           S N S R C T L R E W I I Q G C D F S                                           C L G C T L S Q W R K R T R C D T H                                           Y R G C S R A Q L L G G E C R K K                                             G R G C T L K Q W K Q G D C G R S                                             V R G G C A L R D W V A G E C F D W T                                         L W R G C T L N G F K S R H C G S P E                                         C T L R S W K H R G C A P                                                     G R G C T R A Q W L A G C C T G H                                             R A G C T L R E F R K G C L A L                                               K R G C T L A E M I R G C N R S N                                             G R G C T L K Q W K Q G D C G R S                                             R W R G C S L A K L K K G A A C G R G                                         R G G C T L R E W R R V R V I N                                               G R G C T L K Q W K Q G D C G R S                                             R Y G C T R H Q W L V G T C V R H                                             ______________________________________                                    

IC₅₀ values for some additional representative peptides are given in thetable below. A variety of methods can be used to evaluate IC₅₀ values.For example, an equilibrium binding ELISA assay, using either MBP-TPO orlacI-peptide tracer, was used to determine whether the peptides inhibitthe binding of TPO to the extracellular domain of the TPO receptor.Typically, the IC₅₀ value were determined using the free peptide. TheICSO value can be determined using the free peptide, which optionallycan be C-terminally amidated, or can be prepared as an ester or othercarboxy amide.

To recreate the exact sequence displayed on the phage, the N-terminaland C-terminal amino acids of the synthetic peptides are often precededby one or two glycine residues. These glycines are not believed to benecessary for binding or activity. Likewise, to mimic the exact sequenceof peptides displayed on polysomes, the C-terminal amino acids of thesynthetic peptides are often preceded by the sequence M A S. Again, thissequence is not believed to be necessary for binding or activity.

IC₅ values are indicated symbolically by the symbols "-", "+", and "++".For examples, those peptides which showed IC₅₀ values in excess of 200μM are indicated with a "-". Those peptides which gave IC₅₀ values ofless than or equal to 200 μM are given a "+", while those which gaveIC₅₀ values of 500 nm or less are indicated with a Those peptides whichgave IC₅₀ values at or near the cutoff point for a particular symbol areindicated with a hybrid designator, e.g., "+/-". Those peptides forwhich IC₅₀ values were not determined are listed as "N.D.". The IC₅₀value for peptides having the structure (SEQ ID NO:15): G G C T L R E WL H G G F C G G was 500 nm or less. (Note the N-terminal and C-terminalamino acids were preceded by two glycines to recreate the exact sequencedisplayed by the phage. These glycines are not believed to be necessaryfor binding or activity.)

                  TABLE 3                                                         ______________________________________                                        SEQ ID NOS 6, 7, 8, 9, 168, 11 & 10,                                          respectively)                                                                 Peptide                  Affinity                                             ______________________________________                                        G G C A D G P T L R E W I S F C G G                                                                   ++                                                    G N A D G P T L R Q W L E G R R P K N                                                                 ++                                                    G G C A D G P T L R E W I S F C G G K                                                                 ++                                                    T I K G P T L R Q W L K S R E H T S                                                                   ++                                                    G P T L R Q W L         -                                                     L A I E G P T L R Q W L H G N G R D T                                                                 ++                                                    S I E G P T L R E W L T S R T P H S                                                                   ++                                                    ______________________________________                                    

The tables above, especially Table 3, illustrate that a preferred corepeptide comprises a sequence of amino acids (SEQ ID NO:2):

    X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7

where X₁ is C, L, M, P, Q, V; X₂ is F, K, L, N, Q, R, S, T or V; X₃ isC, F, I, L, M, R, S, V or W; X₄ is any of the 20 genetically codedL-amino acids; X₅ is A, D, E, G, K, M, Q, R, S, T, V or Y; X₆ is C, F,G, L, M, S, V, W or Y; and X₇ is C, G, I, K, L, M, N, R or V.

In a preferred embodiment the core peptide comprises a sequence of aminoacids (SEQ ID NO:3):

    X.sub.8 G X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 W X.sub.7

where X₁ is L, M, P, Q, or V; X₂ is F, R, S, or T; X₃ is F, L, V, or W;X₄ is A, K, L, M, R, S, V, or T; X₅ is A, E, G, K, M, Q, R, S, or T; X₇is C, I, K, L, M or V; and each X₈ residue is independently selectedfrom any of the 20 genetically coded L-amino acids, their stereoisomericD-amino acids; and non-natural amino acids. Preferably, each X₈ residueis independently selected from any of the 20 genetically coded L-aminoacids and their stereoisomeric D-amino acids. In a preferred embodiment(SEQ ID NO:4), X₁ is P; X₂ is T; X₃ is L; X₄ is R; X₅ is E or Q; and X₇is I or L.

More preferably, the core peptide comprises a sequence of amino acids(SEQ ID NO:5):

    X.sub.9 X.sub.8 G X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 W X.sub.7

where X₉ is A, C, E, G, I, L , M, P, R, Q, S, T, or V; and X₈ is A, C,D, E, K, L, Q, R, S, T, or V. More preferably, X₉ is A or I; and X₈ isD, E, or K.

Particularly preferred peptides include (SEQ ID NOS: 6-13,respectively): G G C A D G P T L R E W I S F C G G; G N A D G P T L R QW L E G R R P K N; G G C A D G P T L R E W I S F C G G K; T I K G P T LR Q W L K S R E H T S; S I E G P T L R E W L T S R T P H S; L A I E G PT L R Q W L H G N G R D T; C A D G P T L R E W I S F C; and I E G P T LR Q W L A A R A.

In further embodiments of the invention, preferred peptides for use inthis invention include peptides having a core structure comprisingsequence of amino acids: sequence of amino acids (SEQ ID NO:14):

    C X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7

where X₂ is K, L, N, Q, R, S, T or V; X₃ is C, F, I, L, M, R, S or V; X₄is any of the 20 genetically coded L-amino acids; X₅ is A, D, E, G, S, Vor Y; X₆ is C, F, G, L, M, S, V, W or Y; and X₇ is C, G, I, K, L, M, N,R or V. In a more preferred embodiment, X₄ is A, E, G, H, K, L, M, P, Q,R, S, T, or W. In a further embodiment, X₂ is S or T; X₃ is L or R; X₄is R; X₅ is D, E, or G; X₆ is F, L, or W; and X₇ is I, K, L, R, or V.Particularly preferred peptides include (SEQ ID NO:15): G G C T L R E WL H G G F C G G.

In a further embodiment, preferred peptides for use in this inventioninclude peptides having a structure comprising a sequence of amino acids(SEQ ID NO:16):

    X.sub.8 C X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7

where X₂ is F, K, L, N, Q, R, S, T or V; X₃ is C, F, I, L, M, R, S, V orW; X₄ is any of the 20 genetically coded L-amino acids; X₅ is A, D, E,G, K, M, Q, R, S, T, V or Y; X₆ is C, F, G, L, M, S, V, W or Y; X₇ is C,G, I, K, L, M, N, R or V; and X₈ is any of the 20 genetically codedL-amino acids. In some embodiments, X₈ is preferably G, S, Y, or R.

Peptides and peptidomimetics having an IC₅₀ of greater than about 100 mMlack sufficient binding to permit use in either the diagnostic ortherapeutic aspects of this invention. Preferably, for diagnosticpurposes, the peptides and peptidomimetics have an IC₅₀ of about 2 mM orless and, for pharmaceutical purposes, the peptides and peptidomimeticshave an IC₅₀ of about 100 μM or less.

The binding peptide sequence also provides a means to determine theminimum size of a TPOR binding compound of the invention. Using the"encoded synthetic library" (ESL) system or the "very large scaleimmobilized polymer synthesis" system, one can not only determine theminimum size of a peptide with such activity, but one can also make allof the peptides that form the group of peptides that differ from thepreferred motif (or the minimum size of that motif) in one, two, or moreresidues. This collection of peptides can then be screened for abilityto bind to TPO-receptor. These immobilized polymers synthesis systems orother peptide synthesis methods can also be used to synthesizetruncation analogs, deletion analogs, substitution analogs, andcombinations thereof all of the peptide compounds of the invention.

The peptides and peptide mimetics of the present invention were alsoevaluated in a thrombopoietin dependent cell proliferation assay, asdescribed in greater detail in Example 2 below. Cell proliferation ismeasured by techniques known in the art, such as an MTT assay whichcorrelates with ³ H-thymidine incorporation as an indication of cellproliferation (see Mossmann J. Immunol. Methods 65:55 (1983)). Thepeptides tested stimulated proliferation of TPO-R transfected Ba/F3cells in a dose dependent manner as shown in FIG. 1A. These peptideshave no effect on the parental cell line as shown in FIG. 1B.

FIGS. 7 to 9 show the results of a further assay evaluating activity ofthe peptides and peptide mimetics of the invention. In this assay miceare made thrombocytopenic with carboplatin. FIG. 7 depicts typicalresults when Balb/C mice are treated with carboplatin (125 mg/kgintraperitoneally) on Day 0. The dashed lines represent untreatedanimals from three experiments. The solid line representcarboplatin-treated groups in three experiments. The heavy solid linesrepresent historical data. FIG. 8 depicts the effect of carboplatintitration on platelet counts in mice treated with the indicated amountsof carboplatin (in mg/kg, intraperitoneally (ip) on Day 0). FIG. 9depicts amelioration of carboplatin-induced thrombocytopenia on Day 10by peptide AF12513 (513). Carboplatin (CBP; 50-125 mg/kg,intraperitoneally) was administered on Day 0. AF12513 (1 mg/kg, ip) wasgiven on Days.1-9. These results show the peptides of the invention canameliorate thrombocytopenia in a mouse model.

In addition, certain peptides of the present invention can be dimerizedor oligomerized, thereby increasing the affinity and/or activity of thecompounds. To investigate the effect that peptidedimerization/oligomerization has on TPO mimetic potency in cellproliferation assays, a C-terminally biotinylated analog of the peptide(SEQ ID NO:6) G G C A D G P T L R E W I S F C G G was synthesized (SEQID NO:7) (G G C A D G P T L R E W I S F C G G K (Biotin)). The peptidewas preincubated with strentavidin in serum-free HEPES-buffered RPMI ata 4:1 molar ratio. The complex was tested for stimulation of cellproliferation of TPO-R transfected.Ba/F3 cells, as above, alongside freebiotinylated peptide and the unbiotinylated parental peptide. FIG. 2Ashows the results of the assay for the complexed biotinylated peptide(AF 12885 with streptavidin (SA)) for both the transfected and parentalcell lines. FIG. 2B shows the results of the assay for the freebiotinylated peptide (AP 1228S) for both the transfected and parentalcell lines. FIG. 2C shows the results of the assay for streptavidinalone for both the transfected and parental cell lines. These figuresillustrate that the pre-formed complex was approximately 10 times aspotent as the free peptide.

The specificity of the binding and activity of the peptides of theinvention was also examined by studying the cross reactivity of thepeptides for the erythropoieitin receptor (EPO-R). The EPO-R is also amember of the haematopoietin growth factor receptor family, as is TPO-R.The peptides of the invention, as well as TPO, EPO, and a knownEPO-binding peptide, were examined in a cell proliferation assay usingan EPO-dependent cell line. This assay utilized FDCP-1, a growth factordependent murine multi-potential primitive haematopoietic progenitorcell line (see, e.g., Dexter et al. J. Exp. Med. 152:1036-1047 (1981))as the parental cell line. This cell line can proliferate, but notdifferentiate when supplemented with WEHI-3-conditioned media (a mediumthat contains IL-3, ATCC number T1B68). The parental cell line istransfected with human or murine EPO-R to produce the FDCP-1-EPO-R cellline. These transfected cell lines can proliferate, but notdifferentiate in the presence of human or murine EPO.

The cells were grown to half stationary density in the presence of thenecessary growth factors. The cells are then washed in PBS and starvedfor 16-24 hours in whole media without the growth factors. Afterdetermining the viability of the cells, stock solutions (in whole mediawithout the growth factors) are made to give about 10⁵ cells per 50microliters. Serial dilutions of the compounds (typically, the freesolution phase peptide as opposed to a phage-bound or other bound orimmobilized peptide) to be tested are made in 96-well tissue cultureplates for a final volume of 50 microliters per well. Cells (50microliters) are added to each well and the cells are incubated for24-48 hours, at which point the negative controls should die or bequiescent.

Cell proliferation is then measured by techniques known in the art, suchas an MTT assay.

FIGS. 3A-G show the results of a series of control experiments showingthe activity of TPO, the peptides of the present invention, EPO, andEPO-R binding peptides in a cell proliferation assay using either theTPO-R transfected Ba/F3 cell line and its corresponding parental line,or an EPO-dependent cell line and its corresponding parental line, FIG.3A depicts the results for TPO in the cell proliferation assay using theTPO-R transfected Ba/F3 cell line and its corresponding parental line.FIG. 3B depicts the results for EPO in the cell proliferation assayusing the TPO-R transfected Ba/F3 cell line and its correspondingparental line. FIG. 3C depicts the results for complexed biotinylatedpeptide (AF 12285 with streptavidin (SA)) and a complexed form of abiotinylated EPO-R binding peptide (AF 11505 with SA) in the TPO-Rtransfected Ba/F3 cell line. The results for the corresponding parentalcell line are shown in FIG. 3D. FIG. 3E depicts the results for TPO inthe cell proliferation assay using the EPO-dependent cell line. FIG. 3Fdepicts the results for EPO in the cell proliferation assay using theEPO-dependent cell line. FIG. 3G depicts the results for complexedbiotinylated peptide (AF 1228S with streptavidin (SA)) and the complexedform of a biotinylated EPO-R binding peptide (AF 11505 with SA) in theEPO-dependent cell line. These results show that the peptides of theinvention bind and activate the TPO-R with a high degree of specificity.

IV. PREPARATION OF PEPTIDES AND PEPTIDE MIMETICS

A. SOLID PHASE SYNTHESIS

The peptides of the invention can be prepared by classical methods knownin the art, for example, by using standard solid phase techniques. Thestandard methods include exclusive solid phase synthesis, partial solidphase synthesis methods, fragment condensation, classical solutionsynthesis, and even by recombinant DNA technology. See, e.g., MerrifieldJ. Am. Chem. Soc. 85:2149 (1963), incorporated herein by reference. Onsolid phase, the synthesis is typically commenced from the C-terminalend of the peptide using an alpha-amino protected resin. A suitablestarting material can be prepared, for instance, by attaching therequired alpha-amino acid to a chloromethylated resin, a hydroxymethylresin, or a benzhydrylamine resin. One such chloromethylated resin issold under the tradename BIO-BEADS SX-1 by Bio Rad Laboratories,Richmond, CA, and the preparation of the hydroxymethyl resin isdescribed by Bodonszky et al. Chem. Ind. (London) 38:1597 (1966). Thebenzhydrylamine (BHA) resin has been described by Pietta and MarshallChem. Commn. 650 (1970) and is commercially available from BeckmanInstruments, Inc., Palo Alto, Calif., in the hydrochloride form.

Thus, the compounds of the invention can be prepared by coupling analpha-amino protected amino acid to the chloromethylated resin with theaid of, for example, cesium bicarbonate catalyst, according to themethod described by Gisin Helv. Chim. Acta. 56:1467 (1973). After theinitial coupling, the alpha-amino protecting group is removed by achoice of reagents including trifluoroacetic acid (TFA) or hydrochloricacid (HCl) solutions in organic solvents at room temperature.

The alpha-amino protecting groups are those known to be useful in theart of stepwise synthesis of peptides. Included are acyl type protectinggroups (e.g. Formyl, trifluoroacetyl, acetyl), aromatic urethane typeprotecting groups (e.g. benzyloxycarboyl (Cbz) and substituted Cbz),aliphatic urethane protecting groups (e.g. t-butyloxycarbonyl (Boc),isopropyloxycarbonyl, cyclohexyloxycarbonyl) and alkyl type protectinggroups (e.g. benzyl, triphenylmethyl). Boc and Fmoc are preferredprotecting groups. The side chain protecting group remains intact duringcoupling and is not split off during the deprotection of theamino-terminus protecting group or during coupling. The side chainprotecting group must be removable upon the completion of the synthesisof the final peptide and under reaction conditions that will not alterthe target peptide.

The side chain protecting groups for Tyr include tetrahydropyranyl,tert-butyl, trityl, benzyl, Cbz, Z-Br-Cbz, and 2,5-dichlorobenzyl. Theside chain protecting groups for Asp include benzyl, 2,6-dichlorobenzyl,methyl, ethyl, and cyclohexyl. The side chain protecting groups for Thrand Ser include acetyl, benzoyl, trityl, tetrahydropyranyl, benzyl,2,6-dichlorobenzyl, and Cbz. The side chain protecting group for Thr andSer is benzyl. The side chain protecting groups for Arg include nitro,Tosyl (Tos), Cbz, adamantyloxycarbonyl mesitoylsulfonyl (Mts), or Boc.The side chain protecting groups for Lys include Cbz,2-chlorobenzyloxycarbonyl (2-Cl-Cbz), 2-bromobenzyloxycarbonyl(2-BrCbz), Tos, or Boc.

After removal of the alpha-amino protecting group, the remainingprotected amino acids are coupled stepwise in the desired order. Anexcess of each protected amino acid is generally used with anappropriate carboxyl group activator such as dicyclohexylcarbodiimide(DCC) in solution, for example, in methylene chloride (CH₂ Cl₂),dimethyl formamide (DMF) mixtures.

After the desired amino acid sequence has been completed, the desiredpeptide is decoupled from the resin support by treatment with a reagentsuch as trifluoroacetic acid or hydrogen fluoride (HF), which not onlycleaves the peptide from the resin, but also cleaves all remaining sidechain protecting groups. When the chloromethylated resin is used,hydrogen fluoride treatment results in the formation of the free peptideacids. When the benzhydrylamine resin is used, hydrogen fluoridetreatment results directly in the free peptide amide. Alternatively,when the chloromethylated resin is employed, the side chain protectedpeptide can be decoupled by treatment of the peptide resin with ammoniato give the desired side chain protected amide or with an alkylamine togive a side chain protected alkylamide or dialkylamide. Side chainprotection is then removed in the usual fashion by treatment withhydrogen fluoride to give the free amides, alkylamides, ordialkylamides.

These solid phase peptide synthesis procedures are well known in the artand further described in Stewart Solid Phase Peptide Syntheses (Freemanand Co., San Francisco, (1969)).

Using the "encoded synthetic library" or "very large scale immobilizedpolymer synthesis" system described in U.S. patent application Ser. Nos.07/492,462, filed Mar. 7, 1990; 07/624,120, filed Dec. 6, 1990; and07/805,727, filed Dec. 6, 1991; one can not only determine the minimumsize of a peptide with such activity, one can also make all of thepeptides that form the group of peptides that differ from the preferredmotif (or the minimum size of that motif) in one, two, or more residues.This collection of peptides can then be screened for ability to bind toTPO-R. This immobilized polymer synthesis system or other peptidesynthesis methods can also be used to synthesize truncation analogs anddeletion analogs and combinationc of truncation and deletion analogs ofall of the peptide compounds of the invention.

B. SYNTHETIC AMINO ACIDS

These procedures can also be used to synthesize peptides in which aminoacids other than the 20 naturally occurring, genetically encoded aminoacids are substituted at one, two, or more positions of any of thecompounds of the invention. For instance, naphthylalanine can besubstituted for tryptophan, facilitating synthesis. Other syntheticamino acids that can be substituted into the peptides of the presentinvention include L-hydroxypropyl, L-3, 4-dihydroxyphenylalanyl, d aminoacids such as L-d-hydroxylysyl and D-d-methylalanyl, L-a-methylalanyl, bamino acids, and isoquinolyl. D amino acids and non-naturally occurringsynthetic amino acids can also be incorporated into the peptides of thepresent invention.

One can replace the-naturally occurring side chains of the 20genetically encoded amino acids (or D amino acids) with other sidechains,, for instance with groups such as alkyl, lower alkyl, cyclic 4-,5-, 6-, to 7-membered alkyl, amide, amide lower alkyl, amide di(loweralkyl), lower alkoxy, hydroxy, carboxy and the lower ester derivativesthereof, and with 4-, 5-, 6-, to 7-membered hetereocyclic. Inparticular, proline analogs in which the ring size of the pralineresidue is changed from 5 members to 4, 6, or 7 members can be employed.Cyclic groups can be saturated or unsaturated, and if unsaturated, canbe aromatic or non-aromatic.

Cyclic groups can be saturated or unsaturated, and if unsaturated, canbe aromatic or non-aromatic. Heterocyclic groups preferably contain oneor more nitrogen, oxygen, and/or sulphur heteroatoms. Examples of suchgroups include the furazanyl, furyl, imidazolidinyl, imidazolyl,imidazolinyl, isothiazolyl, isoxazolyl, morpholinyl (e.g. morpholino),oxazolyl, piperazinyl (e.g. 1-piperazinyl), piperidyl (e.g 1-piperidyl,piperidino), pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl,pyridazinyl, pyridyl, pyrimidinyl, pyrrolidinyl (e.g. 1-pyrrolidinyl),pyrrolinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, thiomorpholinyl(e.g. thiomorpholino), and triazolyl. These heterocyclic groups can besubstituted or unsubstituted. Where a group is substituted, thesubstituent can be alkyl, alkoxy, halogen, oxygen, or substituted orunsubstituted phenyl.

One can also readily modify the peptides of the instant invention byphosphorylation, and other methods for making peptide derivatives of thecompounds of the present invention are described in Hruby et al.⁴² Thus,the peptide compounds of the invention also serve as a basis to preparepeptide mimetics with similar biological activity.

The peptide compounds of the invention, including peptidomimetics, canbe covalently modified to one or more of a variety of nonproteinaceouspolymers, e.g., polyethylene glycol, polypropylene glycol, orpolyoxyalkenes, in the manner set forth in U.S. Pat. No. 4,640,835; U.S.Pat. No. 4,496,689; U.S. Pat. No. 4,301,144; U.S. Pat. No. 4,670,417;U.S. Pat. No. 4,791,192; or U.S. Pat. No. 4,179,337, all which-areincorporated by reference in their entirety herein.

C. TERMINAL MODIFICATIONS

Those of skill in the art recognize that a variety of techniques areavailable for constructing peptide mimetics with the same or similardesired biological activity as the corresponding peptide compound butwith more favorable activity than the peptide with respect tosolubility, stability, and susceptibility to hydrolysis and proteolysis.See, for example, Morgan and Gainor Ann. Rep. Med. Chem. 24:243-252(1989). The following describes methods for preparing peptide mimeticsmodified at the N-terminal amino group, the C-terminal carboxyl group,and/or changing ore or more of the amido linkages in the peptide to anon-amido linkage. It being understood that two or more suchmodifications can be coupled in one peptide mimetic structure (e.g.,modification at the C-terminal carboxyl group and inclusion of a --CH₂-carbamate linkage between two amino acids in the peptide).

1. N-TERMINAL MODIFICATIONS

The peptides typically are synthesized as the free acid but, as notedabove, could be readily prepared as the amide or ester. One can alsomodify the amino and/or carboxy terminus of the peptide compounds of theinvention to produce other compounds of the invention. Amino terminusmodifications include methylating (i.e., --NHCH₃ or --NH(CH₃)₂),acetylating, adding a carbobenzoyl grout, or blocking the amino terminuswith any blocking group containing a carboxylate functionality definedby RCOO--, where R is selected from the group consisting of naphthyl,acridinyl, steroidyl, and similar groups. Carboxy terminus modificationsinclude replacing the free acid with a carboxamide group or forming acyclic lactam at the carboxy terminus to introduce structuralconstraints.

Amino terminus modifications are as recited above and includealkylating, acetylating, adding a carbobenzoyl group, forming asuccinimide group, etc. Specifically, the N-terminal amino group canthen be reacted as follows:

(a) to form an amide group of the formula RC(O)NH-- where R is asdefined above by reaction with an acid halide [e.g., RC(O)Cl] or acidanhydride. Typically, the reaction can be conducted by contacting aboutequimolar or excess amounts (e.g., about 5 equivalents) of an acidhalide to the peptide in an inert diluent (e.g., dichloromethane)preferably containing an excess (e.g., about 10 equivalents) of atertiary amine, such as diisopropylethylamine, to scavenge the acidgenerated during reaction. Reaction conditions are otherwiseconventional (e.g., room temperature for 30 minutes). Alkylation of theterminal amino to provide for a lower alkyl N-substitution followed byreaction with an acid halide as described above will provide for N-alkylamide group of the formula RC(O)NR--;

(b) to form a succinimide group by reaction with succinic anhydride. Asbefore, an approximately equimolar amount or an excess of succinicanhydride (e.g., about 5 equivalents) can be employed and the aminogroup is converted to the succinimide by methods well known in the artincluding the use of an excess (e.g., ten equivalents) of a tertiaryamine such as diisopropylethylamine in a suitable inert solvent (e.g.,dichloromethane). See, for example, Wollenberg, et al., U.S. Pat. No.4,612,132 which is incorporated herein by reference in its entirety. Itis understood that the succinic group can be substituted with, forexample, C₂ -C₆ alkyl or --SR substituents which are prepared in aconventional manner to provide for substituted succinimide at theN-terminus of the peptide. Such alkyl substituents are prepared byreaction of a lower olefin (C₂ -C₆) with maleic anhydride in the mannerdescribed by Wollenberg, et al., supra. and --SR substituents areprepared by reaction of RSH with maleic anhydride where R is as definedabove;

(c) to form a benzyloxycarbonyl--NH-- or a substitutedbenzyloxycarbonyl--NH-- group by reaction with approximately anequivalent amount or an excess of CBZ-Cl (i.e., benzyloxycarbonylchloride) or a substituted CBZ-Cl in a suitable inert diluent (e.g.,dichloromethane) preferably containing a tertiary amine to scavenge theacid generated during the reaction;

(d) to form a sulfonamide group by reaction with an equivalent amount oran excess (e.g., 5 equivalents) of R-S(O)₂ Cl in a suitable inertdiluent (dichloromethane) to convert the terminal amine into asulfonamide where R is as defined above. Preferably, the inert diluentcontains excess tertiary amine (e.g., ten equivalents) such asdiisopropylethylamine, to scavenge the acid generated during reaction.Reaction conditions are otherwise conventional (e.g., room temperaturefor 30 minutes);

(e) to form a carbamate group by reaction with an equivalent amount oran excess (e.g., 5 equivalents) of R-OC(O)Cl or R-OC(O)OC₆ H₄ -p-NO₂ ina suitable inert diluent (e.g., dichloromethane) to convert the terminalamine into a carbamate where R is as defined above. Preferably, theinert diluent contains an excess (e.g., about 10 equivalents) of atertiary amine, such as diisopropylethylamine, to scavenge any acidgenerated during reaction. Reaction conditions are otherwiseconventional (e.g., room temperature for 30 minutes); and

(f) to form a urea group by reaction with an equivalent amount or anexcess (e.g., 5 equivalents) of R--N=C=O in a suitable inert diluent(e.g., dichloromethane) to convert the terminal amine into a urea (i.e.,RNHC(O)NH--) group where R is as defined above. Preferably, the inertdiluent contains an excess (e.g., about 10 equivalents) of a tertiaryamine, such as diisopropylethylamine. Reaction conditions are otherwiseconventional (e.g., room temperature for about 30 minutes).

2. C-TERMINAL MODIFICATIONS

In preparing peptide mimetics wherein the C-terminal carboxyl group isreplaced by an ester (i.e., --C(O)OR where R is as defined above), theresins used to prepare the peptide acids are employed, and the sidechain protected peptide is cleaved with base and the appropriatealcohol, e.g., methanol. Side chain protecting groups are then removedin the usual fashion by treatment with hydrogen fluoride to obtain thedesired ester.

In preparing peptide mimetics wherein the C-terminal carboxyl group isreplaced by the amide --C(O)NR³ R⁴, a benzhydrylamine resin is used asthe solid support for peptide synthesis. Upon completion of thesynthesis, hydrogen fluoride treatment to release the peptide from thesupport results directly in the free peptide amide (i.e., the C-terminusis --C(O)NH₂). Alternatively, use of the chloromethylated resin duringpeptide synthesis coupled with reaction with ammonia to cleave the sidechain protected peptide from the support yields the free peptide amideand reaction with an alkylamine or a dialkylamine yields a side chainprotected alkylamide or dialkylamide (i.e., the C-terminus is --C(O)NRR¹where R and R¹ are as defined above). Side chain protection is thenremoved in the usual fashion by treatment with hydrogen fluoride to givethe free amides, alkylamides, or dialkylamides.

In another alternative embodiment, the C-terminal carboxyl group or aC-terminal ester can be induced to cyclize by internal displacement ofthe --OH or the ester (--OR) of the carboxyl group or ester respectivelywith the N-terminal amino group to form a cyclic peptide. For example,after synthesis and cleavage to give the peptide acid, the free acid isconverted to an activated ester by an appropriate carboxyl groupactivator such as dicyclohexylcarbodiimide (DCC) in solution, forexample, in methylene chloride (CH₂ Cl₂), dimethyl formamide (DMF)mixtures. The cyclic peptide is then formed by internal displacement ofthe activated ester with the N-terminal amine. Internal cyclization asopposed to polymerization can be enhanced by use of very dilutesolutions. Such methods are well known in the art.

One can also cyclize the peptides of the invention, or incorporate adesamino or descarboxy residue at the terminii or the peptide, so thatthere is no terminal amino or carboxyl group, to decrease susceptibilityto proteases or to restrict the conformation of the peptide. C-terminalfunctional groups of the compounds of the present invention includeamide, amide lower alkyl, amide di(lower alkyl), lower alkoxy, hydroxy,and carboxy, and the lower ester derivatives thereof, and thepharmaceutically acceptable salts thereof.

D. BACKBONE MODIFICATIONS

Other methods for making peptide derivatives of the compounds of thepresent invention are described in Hruby et al. Biochem J.268(2):249-262 (1990), incorporated herein by reference. Thus, thepeptide compounds of the invention also serve as structural models fornon-peptidic compounds with similar biological activity. Those of skillin the art recognize that a variety of techniques are available forconstructing compounds with the same or similar desired biologicalactivity as the lead peptide compound but with more favorable activitythan the lead with respect to solubility, stability, and susceptibilityto hydrolysis and proteolysis. See Morgan and Gainor Ann. Rep. Med.Chem. 24:243-252 (1989), incorporated herein by reference. Thesetechniques include replacing the peptide backbone with a backbonecomposed of phosphonates, amidates, carbamates, sulfonamides, secondaryamines, and N-methylamino acids.

Peptide mimetics wherein one or more of the peptidyl linkages[--C(O)NH--] have been replaced by such linkages as a --CH₂ -carbamatelinkage, a phosphonate linkage, a --CH₂ -sulfonamide linkage, a urealinkage, a secondary amine (--CH₂ NH--) linkage, and an alkylatedpeptidyl linkage [--C(O)NR⁶ -- where R⁶ is lower alkyl] are preparedduring conventional peptide synthesis by merely substituting a suitablyprotected amino acid analogue for the amino acid reagent at theappropriate point during synthesis.

Suitable reagents include, for example, amino acid analogues wherein thecarboxyl group of the amino acid has been replaced with a moietysuitable for forming one of the above linkages. For example, if onedesires to replace a --C(O)NR-- linkage in the peptide with a --CH₂-carbamate linkage (--CH₂ OC(O)NR--), then the carboxyl (--COOH) groupof a suitably protected amino acid is first reduced to the --CH₂ OHgroup which is then converted by conventional methods to a --OC.(O)Clfunctionality or a para-nitrocarbonate --OC(O)O-C₆ H₄ -p-NO₂functionality. Reaction of either of such functional groups with thefree amine or an alkylated amine on the N-terminus of the partiallyfabricated peptide found on the solid support leads to the formation ofa --CH₂ OC(O)NR-- linkage. For a more detailed description of theformation of such --CH₂ -carbamate linkages, see Cho et al Science,261:1303-1305 (1993).

Similarly, replacement of an amido linkage in the peptide with aphosphonate linkage can be achieved in the manner set forth in U.S.patent application Ser. Nos. 07/943,805, 08/081,577, and 08/119,700, thedisclosures of which are incorporated herein by reference in theirentirety.

Replacement of an amido linkage in the peptide with a --CH₂ -sulfonamidelinkage can be achieved by reducing the carboxyl (--COOH) group of asuitably protected amino acid to the --CH₂ OH group and the hydroxylgroup is then converted to a suitable leaving group such as a tosylgroup by conventional methods. Reaction of the tosylated derivativewith, for example, thioacetic acid followed by hydrolysis and oxidativechlorination will provide for the --CH₂ --S(O)₂ Cl functional groupwhich replaces the carboxyl group of the otherwise suitably protectedamino acid. Use of this suitably protected amino acid analogue inpeptide synthesis provides for inclusion of an --CH₂ S(O)₂ NR-- linkagewhich replaces the amido linkage in the peptide thereby providing apeptide mimetic. For a more complete description on the conversion ofthe carboxyl group of the amino acid to a --CH₂ S(O)₂ Cl group, see, forexample, Weinstein, Boris Chemistry & Biochemistry of Amino Acids,Peptides and Proteins Vol. 7, pp. 267-357, Marcel Dekker, Inc., New York(1983) which is incorporated herein by reference.

Replacement of an amido linkage in the peptide with: a urea linkage canbe achieved in the manner set forth in U.S. patent application Ser. No.08/147,805 which application is incorporated herein by reference in itsentirety.

Secondary amine linkages wherein a --CH₂ NH-- linkage replaces the amidolinkage in the peptide can be prepared by employing, for example, asuitably protected dipeptide analogue wherein the carbonyl bond of theamido linkage has been reduced to a CH₂ group by conventional methods.For example, in the case of diglycine, reduction of the amide to theamine will yield after deprotection H₂ NCH₂ CH₂ NHCH₂ COOH which is thenused in N-protected form in the next coupling reaction. The preparationof such analogues by reduction of the carbonyl group of the amidolinkage in the dipeptide is well known in the art.

The suitably protected amino acid analogue is employed in theconventional peptide synthesis in the same manner as would thecorresponding amino acid. For example, typically about 3 equivalents ofthe protected amino acid analogue are employed in this reaction. Aninert organic diluent such as methylene chloride or DMF is employed and,when an acid is generated as a reaction by-product, the reaction solventwill typically contain an excess amount of a tertiary amine to scavengethe acid generated during the reaction. One particularly preferredtertiary amine is diisopropylethylamine which is typically employed inabout 10 fold excess. The reaction results in incorporation into thepeptide mimetic of an amino acid analogue having a non-peptidyl linkage.Such substitution can be repeated as desired such that from zero to allof the amido bonds in the peptide have been replaced by non-amido bonds.

One can also cyclize the peptides of the invention, or incorporate adesamino or descarboxy residue at the terminii of the peptide, so thatthere is no terminal amino or carboxyl group, to decrease susceptibilityto proteases or to restrict the conformation of the peptide. C-terminal,functional groups of the compounds of the present invention includeamide, amide lower alkyl, amide di(lower alkyl), lower alkoxy, hydroxy,and carboxy, and the lower ester derivatives thereof, and thepharmaceutically acceptable salts thereof. Examples of cyclizedcompounds are provided in Tables 4, 5, 6, 8, and 9.

E. DISULFIDE BOND FORMATION

The compounds of the present invention may exist in a cyclized form withan intramolecular disulfide bond between the thiol groups of thecysteines. Alternatively, an intermolecular disulfide bond between thethiol groups of the cysteines can be produced to yield a dimeric (orhigher oligomeric) compound. One or more of the cysteine residues mayalso be substituted with a homocysteine. These intramolecular orintermolecular disulfide derivatives can be represented schematically asshown below: ##STR1## wherein m and n are independently 1 or 2.

Other embodiments of this invention provide for analogs of thesedisulfide derivatives in which one of the sulfurs has been replaced by aCH₂ group or other isostere for sulfur. These analogs can be made via anintramolecular or intermolecular displacement, using methods known inthe art as shown below: ##STR2## wherein p is 1 or 2. One of skill inthe art will readily appreciate that this displacement can also occurusing other homologs of the a-amino-g-butyric acid derivative shownabove and homocysteine.

Alternatively, the amino-terminus of the peptide can be capped with analpha-substituted acetic acid, wherein the alpha substituent is aleaving group, such as an a-haloacetic acid, for example, a-chloroaceticacid, a-bromoacetic acid, or a-iodoacetic acid. The compounds of thepresent invention can be cyclized or dimerized via displacement of theleaving group by the sulfur of the cysteine or homocysteine residue.See, e.g., Barker et al. J. Med. Chem. 35:2040-2048 (1992) and Or et al.J. Org. Chem. 56:3146-3149 (1991), each of which is incorporated hereinby reference. Examples of dimerized compounds are provided in Tables 7,9, and 10.

V. UTILITY

The compounds of the invention are useful in vitro as unique tools forunderstanding the biological role of TPO, including the evaluation ofthe many factors thought to influence, and be influenced by, theproduction of TPO and the receptor binding process. The presentcompounds are also useful in the development of other compounds thatbend to and activate the TPO-R, because the present compounds provideimportant information on the relationship between structure and activitythat should facilitate such development.

The compounds are also useful as competitive binders in assays to screenfor new TPO receptor agonists. In such assay embodiments, the compoundsof the invention can be used without modification or can be modified ina varietv of ways; for example, by labeling, such as covalently ornon-covalently joining a moiety which directly or indirectly provides adetectable signal. In any of these assays, the materials thereto can belabeled either directly or indirectly. Possibilities for direct labelinginclude label groups such as: radiolabels such as ¹²⁵ I, enzymes (U.S.Pat. No. 3,645,090) such as peroxidase and alkaline phosphatase, andfluorescent labels (U.S. Pat. No. 3,940,475) capable of monitoring thechange in fluorescence intensity, wavelength shift, or fluorescencepolarization. Possibilities for indirect labeling include biotinylationof one constituent followed by binding to avidin coupled to one of theabove label groups. The compounds may also include spacers or linkers incases where the compounds are to be attached to a solid support.

Moreover, based on their ability to bind to the TPO receptor, thepeptides of the present invention can be used as reagents for detectingTPO receptors on living cells, fixed cells, in biological fluids, intissue homogenates, in purified, natural biological materials, etc. Forexample, by labelling such peptides, one can identify cells having TPO-Ron their surfaces. In addition, based on their ability to bind the TPOreceptor, the peptides of the present invention can be used in in situstaining, FACS (fluorescence-activated cell sorting), Western blotting,ELISA, etc. In addition, based on their ability to bind to the TPOreceptor, the peptides of the present invention can be used in receptorpurification, or in purifying cells expressing TPO receptors on the cellsurface (or inside permeabilized cells).

The compounds of the present invention can also be utilized ascommercial reagents for various medical research and diagnostic uses.Such uses include but are not limited to: (1) use as a calibrationstandard for quantitating the activities of candidate TPO agonists in avariety of functional assays; (2) use to maintain the proliferation andgrowth of TPO-dependent cell lines; (3) use in structural analysis ofthe TPO-receptor through co-crystallization; (4) use to investigate themechanism of TPO signal transduction/receptor activation; and (5) otherresearch and diagnostic applications wherein the TPO-receptor ispreferably activated or such activation is conveniently calibratedagainst a known quantity of a TPO agonist, and the like.

The compounds of the present invention can be used for the in vitroexpansion of megakaryocytes and their committed progenitors, both inconjunction with additional cytokines or on their own. See, e.g.,DiGiusto et al. PCT Publication No. 95/05843, which is incorporatedherein by reference. Chemotherapy and radiation therapies causethrombocytopenia by killing the rapidly dividing, more mature populationof megakaryocytes. However, these therapeutic treatments can also reducethe number and viability of the immature, less mitotically activemegakaryocyte precursor cells. Thus, amelioration of thethrombocytopenia by TPO or the compounds of the present invention can behastened by infusing patients post chemotherapy or radiation therapywith a population of his or her own cells enriched for megakaryocytesand immature precursors by in vitro culture.

The compounds of the invention can also be administered to warmblooded-animals, including humans, to activate the TPO-R in vivo. Thus,the present invention encompasses methods for therapeutic treatment ofTPO related disorders that comprise administering a compound of theinvention in amounts sufficient to mimic the effect of TPO on TPO-R invivo. For example, the peptides and compounds of the invention can beadministered to treat a variety of hematological disorders, includingbut not limited to platelet disorders and thrombocytopenia, particularlywhen associated with bone marrow transfusions, radiation therapy, andchemotherapy.

In some embodiments of the invention, TPO antagonists are preferablyfirst administered to patients undergoing chemotherapy or radiationtherapy, followed by administration of the tpo agonists of theinvention.

The activity of the compounds of the present invention can be evaluatedeither in vitro or in vivo in one of the numerous models described inMcDonald Am. J. of Pediatric Hematology/Oncology 14:8-21 (1992) , whichis incorporated herein by reference.

According to one embodiment, the compositions of the present inventionare useful for treating thrombocytopenia associated with bone marrowtransfusions, radiation therapy, or chemotherapy. The compoundstypically will be administered prophylactically prior to chemotherapy,radiation therapy, or bone marrow transplant or after such exposure.

Accordingly, the present invention also provides pharmaceuticalcompositions comprising, as an active ingredient, at least one of thepeptides or peptide mimetics of the invention in association with apharmaceutical carrier or diluent. The compounds of this invention canbe administered by oral, pulmonary, parental (intramuscular,intraperitoneal, intravenous (IV) or subcutaneous injection), inhalation(via a fine powder formulation), transdermal, nasal, vaginal, rectal, orsublingual routes of administration and can be formulated in dosageforms appropriate for each route of administration. See, e.g., Bernsteinet al. PCT Patent Publication No. WO 93/25221; Pitt et al. PCT PatentPublication No. WO 94/17784; and Pitt et al. European Patent Application613,683, each of which is incorporated herein by reference.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is admixed with at least one inert pharmaceutically acceptablecarrier such as sucrose, lactose, or starch. Such dosage forms can alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., lubricating agents such as magnesium stearate. In thecase of capsules, tablets, and pills, the dosage forms may also comprisebuffering agents. Tablets and pills can additionally be prepared withenteric coatings.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, with the elixirscontaining inert diluents commonly used in the art, such as water.Besides such inert diluents, compositions can also include adjuvants,such as wetting agents, emulsifying and suspending agents, andsweetening, flavoring, and perfuming agents.

Preparations according to this invention for parental administrationinclude sterile aqueous or non-aqueous solutions, suspensions, oremulsions. Examples of non-aqueous solvents or vehicles are propyleneglycol, polyethylene glycol, vegetable oils, such as olive oil and cornoil, gelatin, and injectable organic esters such as ethyl oleate. Suchdosage forms may also contain adjuvants such as preserving, wetting,emulsifying, and dispersing agents. They may be sterilized by, forexample, filtration through a bacteria retaining filter, byincorporating sterilizing agents into the compositions, by irradiatingthe compositions, or by heating the compositions. They can also bemanufactured using sterile water, or some other sterile injectablemedium, immediately before use.

Compositions for rectal or vaginal administration are preferablysuppositories which may contain, in addition to the active substance,excipients such as cocoa butter or a suppository wax. Compositions fornasal or sublingual administration are also prepared with standardexcipients well known in the art.

The compositions containing the compounds can be administered forprophylactic and/or therapeutic treatments. In therapeutic applications,compositions are administered to a patient already suffering from adisease, as described above, in an amount sufficient to cure or at leastpartially arrest the symptoms of the disease and its complications. Anamount adequate to accomplish this is defined as "therapeuticallyeffective dose". Amounts effective for this use will depend on theseverity of the disease and the weight and general state of the patient.

The compositions of the invention can also be microencapsulated by, forexample, the method of Tice and Bibi (in Treatise on Controlled DrugDelivery, ed. A. Kydonieus, Marcel Dekker, N.Y. (1992), pp. 315-339).

In prophylactic applications, compositions containing the compounds ofthe invention are administered to a patient susceptible to-or otherwiseat risk of a particular disease. Such an amount is defined to be a"prophylactically effective dose". In this use, the precise amountsagain depend on the patient's state of health and weight.

The quantities of the TPO agonist necessary for effective therapy willdepend upon many different factors, including means of administration,target site, physiological state of the patient, and other medicantsadministered. Thus, treatment dosages should be titrated to optimizesafety and efficacy. Typically, dosages used in vitro may provide usefulguidance in the amounts useful for in situ administration of thesereagents. Animal testing of effective doses for treatment of particulardisorders will provide further predictive indication of human dosage.Various considerations are described, e.g., in Gilman et al. (eds),Goodman and Gilman's: The Pharmacological Basis of Therapeutics, 8thed., Pergamon Press (1990); and Remington's Pharmaceutical Sciences, 7thed., Mack Publishing Co., Easton, Pa. (1985); each of which is herebyincorporated by reference.

The peptides and peptide mimetics of this invention are effective intreating TPO mediated conditions when administered at a dosage range offrom about 0.001 mg to about 10 mg/kg of body weight per day. Thespecific dose employed is regulated by the particular condition beingtreated, the route of administration as well as by the judgement of theattending clinician depending upon factors such as the severity of thecondition, the age and general condition of the patient, and the like.

Although only preferred embodiments of the invention are specificallydescribed above, it will be appreciated that modifications andvariations of the invention are possible without departing from thespirit and intended scope of the invention.

EXAMPLE 1 SOLID PHASE PEPTIDE SYNTHESIS

Various peptides of the invention were synthesized using the Merrifieldsolid phase synthesis techniques (See Steward and Young, Solid PhasePeptide Synthesis, 2d. edition, Pierce Chemical, Rockford, Ill. (1984)and Merrifield J. Am. Chem. Soc. 85:2149 (1963)) on a Milligen/Biosearch9600 automated instrument or an Applied Biosystems Inc. Model 431Apeptide synthesizer. The peptides were assembled using standardprotocols of the Applied Biosystems Inc. System Software version 1.01.Each coupling was performed for one-two hours with BOP (benzotriazolylN-oxtrisdimethylaminophosphonium hexafluorophosphate) and HOBt(1-hydroxybenzotriazole).

The resin used was HMP resin or PAL (Milligen/Biosearch), which is across-linked polystyrene resin with5-(4'-Fmoc-aminomethyl-3,5'-dimethyoxyphenoxy) valeric acid as a linker.Use of PAL resin results in a carboxyl terminal amide functionality uponcleavage of the peptide from the resin. Upon cleavage, the HMP resinproduces a carboxylic acid moiety at the C-terminus of the finalproduct. Most reagents, resins, and protected amino acids (free or onthe resin) were purchased from Millipore or Applied Biosystems Inc.

The Fmoc group was used for amino protection during the couplingprocedure. Primary amine protection on amino acids was achieved withFmoc and side chain protection groups were t-butyl for serine, tyrosine,asparagine, glutamic acid, and threonine; trityl for glutamine; Pmc(2,2,5,7,8-pentamethylchroma sulfonate) for arginine;N-t-butyloxycarbonyl for tryptophan; N-trityl for histidine andglutamine; and S-trityl for cysteine.

Removal of the peptides from the resin and simultaneous deprotection ofthe side chain functions were achieved by treatment with reagent K orslight modifications of it. Alternatively, in the synthesis of thosepeptides, with an amidated carboxyl terminus, the fully assembledpeptide was cleaved with a mixture of 90% trifluoroacetic acid, 5%ethanedithiol, and 5% water, initially at 4° C., and graduallyincreasing to room temperature. The deprotected peptides wereprecipitated with diethyl ether. In all cases, purification was bypreparative, reverse-phase, high performance liquid chromatography on aC₁₈ bonded silica gel column with a gradient of acetonitrile/water in0.1% trifluoroacetic acid. The homogeneous peptides were characterizedby Fast Atom Bombardment mass spectrometry or electrospray massspectrometry and amino acid analysis when applicable.

EXAMPLE 2 BIOASSAYS

Bioactivity of the peptides can be measured using a thrombopoietindependent cell proliferation assay. Murine IL-3 dependent Ba/F3 cellswere transfected with full length human TPO-R. In the absence of IL-3(WEHI-3 conditioned media), these cells are dependent on TPO forproliferation. The parental, untransfected cell line does not respond tohuman TPO, but remains IL-3 dependent.

Bioassays have been performed on both of the above cell lines usingsynthetic peptides derived from library screening. The cells were grownin complete RPMI-10 media, containing 10% WEHI-3 conditioned media, thenwashed twice in PBS, resuspended in media which lacked WEHI-3conditioned media, and added to wells containing dilutions of peptide orTPO at 2×10⁴ cells/well. The cells were incubated for 48 hours at 37° C.in a humidified 5% CO₂ atmosphere and metabolic activity was assayed bythe reduction of MTT to formazan, with absorbance at 570 nM measured onan ELISA plate reader. The peptides tested stimulated proliferation ofTPO-R transfected Ba/F3 cells in a dose dependent manner as shown inFIG. 1. These peptides have no effect on the parental cell line.

EXAMPLE 3 BINDING AFFINITY

Binding affinities of chemically synthesized peptides for TPO-R weremeasured in a competition binding assay. The wells of a microtiter platewere coated with 1 mg streptavidin, blocked with PBS/1% BSA, followed by50 ng of biotinylated anti-receptor immobilizing antibody (Ab179). Thewells were then treated with a 1:10 dilution of soluble TPO-R harvest.Various concentrations of peptide or peptide mimetic were mixed with aconstant amount of a truncated form of TPO consisting of residues 1-156fused to the C-terminus of maltose binding protein (MBP-TPO₁₅₆) . Thepeptide MBP-TPO₁₅₆ mixtures were added to the TPO-R coated wells,incubated for 2 hours at 4° C. and then washed with PBS. The amount ofMBP-TPO₁₅₆ that was bound at equilibrium was measured by adding a rabbitanti-sera directed against MBP, followed by alkaline phosphataseconjugated goat anti-rabbit IgG. The amount of alkaline phosphatase ineach well was then determined using standard methods.

The assay is conducted over a range of peptide concentrations and theresults are graphed such that the y axis represents the amount of boundMBP-TPO₁₅₆ and the x axis represents the concentration of peptide orpeptide mimetic. One can then determine the concentration at which thepeptide or peptide mimetic will reduce by 50% (IC₅₀) the amount ofMBP-TPO₁₅₆ bound to immobilized TPO-R. The dissociation constant (Kd)for the peptide should be similar to the measured IC₅₀ so using theassay conditions described above.

EXAMPLE 4 "PEPTIDES ON PLASMIDS"

The pJS142 vector is used for library construction and is shown in FIG.4. Three oligonucleotide sequences (SEQ ID NOS:169-171, respectively)are needed for library construction: ON-829 (5' ACC ACC TCC GG); ON-830(5' TTA CTT AGT TA) and a library specific oligonucleotide of interest(5' GA GGT GGT {NNK}_(n) TAA CTA AGT AAA GC), where {NNK}_(n) denotes arandom region of the desired length and sequence. The oligonucleotidescan be 5' phosphorylated chemically during synthesis or afterpurification with polynucleotide kinase. They are then annealed at a1:1:1 molar ratio and ligated to the vector.

The strain of E. coli which is preferably used for canning has thegenotype: Δ(srl-recA) endal nupG lon-11 sulAl hsdR17 A(ompT-fepC)266ΔclpA319::kan ΔlacI lac ZU118 which can be prepared from an E. colistrain from the E. coli Genetic Stock Center at Yale University (E. colib/r, stock center designation CGSC:6573) with genotype lon-11 sulA1. Theabove E. coli strain is prepared for use in electroporation as describedby Dower et al. Nucleic Acids Res. 16:6127 (1988), except that 10%glycerol is used for all wash steps. The cells are tested for efficiencyusing 1 pg of a Bluescript plasmid (Stratagene). These cells are usedfor growth of the original library and for amplification of the enrichedpopulation after each round of panning.

Peptides on plasmids are released from cells for panning by gentleenzymatic digestion of the cell wall using lysozyme. After pelleting ofthe cell debris, the crude lysate can be used directly on mostreceptors. If some additional purification of the plasmid complexes isneeded, a gel filtration column can be used to remove many of the lowmolecular weight contaminants in the crude lysate.

Panning is carried out in a buffer (HEKL) of a lower salt concentrationthan most physiological buffers. The panning can be conducted inmicrotiter wells with a receptor immobilized on a nonblocking monoclonalantibody (MAb) or by panning on beads or on columns. More specifically,in the first round of panning, 24 wells, each coated with receptor, canbe used. For the second round, six wells coated with receptor (PANsample) and 6 wells without receptor (NC sample) are typically used.Comparison of the number of plasmids in these two samples can give anindication of whether receptor specific clones are being enriched bypanning. "Enrichment" is defined as the ratio of PAN transformants tothose recovered from the NC sample. Enrichment of 10 fold is usually anindication that receptor specific clones are present.

In later rounds of panning, it is useful to reduce. the input of lysateinto the wells to lower nonspecific background binding of the plasmidcomplexes. In round 2, usually 100 μl of lysate per well is used. Inround 3, 100 μl of lysate per well diluted with 1/10 in HEKL/BSA isused. For further rounds of panning, typically an input of plasmidtransforming units of at least 1000 fold above the estimated remainingdiversity is used.

The binding properties of the peptides encoded by individual clones aretypically examined after 3, 4, or 5 rounds of panning, depending on theenrichment numbers observed. Typically, an ELISA that detects receptorspecific binding by LacI-peptide fusion proteins is used. LacI isnormally a tetramer and the minimum functional DNA binding species is adimer. The peptides are thus displayed multivalently on the fusionprotein. Assuming that a sufficient density of receptor can beimmobilized in wells, the peptides fused to LacI will bind to thesurface in a cooperative, multivalent fashion. This cooperative bindingpermits the detection of binding events of low intrinsic affinity. Thesensitivity of this assay is an advantage in that initial hits of lowaffinity can be easily identified, but is a disadvantage in that thesignal in the ELISA is not correlated with the intrinsic affinity of thepeptides. Fusion of the peptides to maltose binding protein (MBP) asdescribed below permits testing in an ELISA format where signal strengthis better correlated with affinity. See FIGS. 5A-B.

DNA from clones of interest can be prepared in double stranded formusing any standard miniprep procedure. The coding sequences ofinteresting single clones or populations of clones can be transferred tovectors that fuse those sequences in frame with the gene encoding MBP, aprotein that generally occurs as a monomer in solution. The cloning of alibrary into pJS142 creates a BspEI restriction site near the beginningof the random coding region of the library.

Digestion with BspEI and ScaI allows the purification of a .sup.˜ 900 bpDNA fragment that can be subcloned into one of two vectors, pELM3(cytoplasmic) or pELM15 (periplasmic), which are simple modifications ofthe pMALc2 and PMALp2 vectors, respectively, available commercially fromNew England Biolabs. See FIGS. 5A-B. Digestion of pELM3 and pELM15 withAgeI and ScaI allows efficient cloning of the BspEI-ScaI fragment fromthe pJS142 library. The BspEI and AgeI ends are compatible for ligation.In addition, correct ligation of the ScaI sites is essential to recreatea functional bla (Amp resistance) gene, thus lowering the level ofbackground clones from undesired ligation events. Expression of the tacpromoter-driven MBP-peptide fusions can then be induced with IPTG.

Lysates for the LacI or MBP ELISAs are prepared from individual clonesby lysing cells using lysozyme and removing insoluble cell debris bycentrifugation. The lysates are then added to wells containingimmobilized receptor and to control wells without receptor. Binding bythe LacI or MBP peptide fusions is detected by incubation with a rabbitpolyclonal antiserum directed against either LacI or MBP followed byincubation with alkaline phosphatase labeled goat anti rabbit secondantibody. The bound alkaline phosphatase is detected with p-nitrophenylphosphate chromagenic substrate.

EXAMPLE 5 "HEADPIECE DIMER" SYSTEM

A variant of the LacI peptides-on-plasmids technique utilizes a DNAbinding protein called "headpiece dimer". DNA binding by the E. coli lacrepressor is mediated by the approximately 60 amino acid "headpiece"domain. The dimer of the headpiece domains that binds to the lacoperator is normally formed by association of the much largerapproximately 300 amino acid C-terminal domain. The "headpiece dimer"system utilizes headpiece dimer molecules containing two headpiecesconnected via short peptide linker. These proteins bind DNA withsufficient stability to allow association of a peptide epitope displayedat the C-terminus of the headpiece dimer with the plasmid encoding thatpeptide.

The random peptides are fused to the C-terminus of the headpiece dimer,which binds to the plasmid that encoded it to make a peptide-headpiecedimer-plasmid complex that can be screened by panning. The headpiecedimer peptides-on-plasmids system allows greater selectivity for highaffinity ligands than the LacI system. Thus, the headpiece dimer systemis useful for making mutagenesis libraries based on initial low-affinityhits, and selecting higher affinity variants of those initial sequences.

The libraries are constructed as with peptides on plasmids usingheadpiece dimer vector pCMG14 (see FIGS. 6A-C). The presence of the lacoperator is not required for plasmid binding by the headpiece dimerprotein. The libraries were introduced into bacterial strain comprisingE. coli (lon-11 sulAl hsdR17 (ompT-fepC) ΔclpA319::kan ΔlacI lac ZU118Δ(srl-recA) 306::Tn10 and amplified under conditions of basal (A)promoter induction. Panning of headpiece dimer libraries is carried outby similar procedures to those used for LacI libraries, except that HEKbuffer is used instead of HEKL buffer and elution of plasmids from thewells is performed with aqueous phenol instead of with IPTG. Sequencesfrom headpiece dimer panning are often characterized after transfer tothe MBP vector so that they can be tested in the affinity sensitive MBPELISA and also so that populations of clones can be screened by colonylifts with labeled receptor.

EXAMPLE 6

In this example cyclized compounds were subjected to three assays.First, IC₅₀ valves were obtained as described above. Additionally, anMTT cell proliferation assay as described above was performed tocalculate EC₅₀ values. Finally, a microphysiometer (Molecular DevicesCorp.) assay was performed. Basically, in this assay the rate ofacidification of the extracellular medium in response to TPO receptorstimulation by the compounds of the invention was determined. The rangesfor EC₅₀ are symbolically indicated as for IC₅₀ described above. Theresults are summarized in Table 4.

                                      TABLE 4                                     __________________________________________________________________________                                    EC50 (nM)                                                                           EC50 (nM)                               Structure                       Proliferation                                                                       Microonys.                                                                          IC50 (nM)                         __________________________________________________________________________     ##STR3##                       ++    ++    ++                                 ##STR4##                       ++    ++    ++                                 ##STR5##                       ++    ++    ND                                 ##STR6##                       +     +-    +                                  ##STR7##                       +     +-    ND                                 ##STR8##                       +-    +     ++                                 ##STR9##                       +     +     ++                                 ##STR10##                      +     +     ++                                 ##STR11##                      +     +     ++                                 ##STR12##                      +     +     +-                                 ##STR13##                      ++    +     ++                                 ##STR14##                      +     +-    ND                                 ##STR15##                      +     +     ND                                 ##STR16##                      ++    +     ND                                 ##STR17##                      ++    +-    ND                                 ##STR18##                      +-    +-    +-                                 ##STR19##                      +-    +-    ND                                __________________________________________________________________________

EXAMPLE 7

In this example amino acid substitutes at positions D, E, I, S, or F inthe cyclized compound (SEQ ID NO:12) ##STR20## were assayed for EC₅₀ andIC₅₀ values as described above. Microphysiometer results are given inparentheses. The results are summarized in Table 5 below.

                  TABLE 5                                                         ______________________________________                                        (SEQ ID NO:12)                                                                 ##STR21##                                                                                    EC50 (nM)                                                     Substitution    Cell Prolif.                                                                            IC50 (nM)                                           ______________________________________                                        E-Q             ++(+)     ++                                                  D-A             +(+)      ++                                                  I-A             +-(+)     +                                                   S-A             ++(++)    ++                                                  S-D-Ala         +         +-                                                  S-Sar           +-        ++                                                  S-Aib           ++(+)     ++                                                  S-D-Ser         ++        ++                                                  S-Nva           ++(++)    ++                                                  S-Abu           ++        ++                                                  S-(N-Me-Ala)    +         +                                                   S-(N-Me-Val)    +         +-                                                  S-(N-Me-Ala)*   +         +-                                                  S-(Nor-Leu)     ++        ++                                                  S-(t-Bu-Gly)    +         ++                                                  S-[N-Me-Ser(Bzl)]         +                                                   S-(Homoser)     ND        ND                                                  S-(N-Me-Leu)    +         ND                                                  F-A             +(+)      ++                                                  F-D-Ala         +         ++                                                  F-D-Phe         +         ++                                                  F-Homo-Phe      ++(++)    ++                                                  F-CHA           ++(++)    ++                                                  F-Thi           ++        ++                                                  F-(Ser(Bzl))    ++        ++                                                  F-(N-Me-Ala)    +-        +-                                                  F-(Phenyigly)   ++(++)    ++                                                  F-(Pyridylala)  ++        ++                                                  F-(p-Nitrophe)  ++(++)    ++                                                  F-(3,4-di-Cl-Phe)                                                                             ++(+)     ++                                                  F-(p-Cl-Phe)    ++        ++                                                  F-(2-Nal)       ++(++)    ++                                                  F-(1-Nal)       ++        ++                                                  F-(DiPh-Ala)    ++        ++                                                  F-(N-Me-Phe)    ++        ND                                                  S,F-Ava (thioether)                                                                           +-        ++                                                  S,F-Ava (cys-cys)                                                                             +         ++                                                  S,F-Ava         +-        ++                                                  AD-deletion     +-(+)     ND                                                  ADG-deletion    (+)       +                                                   ______________________________________                                         ##STR22##                                                                     -                                                                        

EXAMPLE 8

In this example, amino acid substitutions in the compound (SEQ IDNO:173) ##STR23## were evaluated at positions D, S, or F as indicated inTable 6 below. EC₅₀ and IC₅₀ values were calculated as described above.Microphysiometer results are in parentheses.

                  TABLE 6                                                         ______________________________________                                        (SEQ ID NO:173)                                                                ##STR24##                                                                                     EC50 (nM)                                                    Substitution     Cell Prolif.                                                                            IC50 (nM)                                          ______________________________________                                        D-E              (+)       ND                                                 free acid form   ++(+)     ND                                                 C-term. Gly addition                                                                           ++        ++                                                 S-Abu            ++(++)    ND                                                 F-DiPh-Ala       (++)      ++                                                 S,F-Abu, DiPh-Ala                                                                              +-(+)     ++                                                 ______________________________________                                    

EXAMPLE 9

In this example EC₅₀ and IC₅₀ values were calculated as described abovefor the dimer compounds listed in Table 7 below. The cyclized monomer(SEQ ID NO:12) ##STR25## is included as a comparison.

The compounds of Table 8 were inactive at the maximum concentrationtested of 10 μm.

In Table 9, EC₅₀ and IC₅₀ values determined as described above forcyclized and dimerized variants of (SEQ ID NO:193) I E G P T L R Q W L AA R A are compared.

In Table 10, truncations of the dimer (SEQ ID NOS: 17-18, respectively)##STR26## are compared. EC₅₀ and IC₅₀ values were calculated asdescribed above. Microphysiometer results are given in parentheses.

                                      TABLE 7                                     __________________________________________________________________________                               EC50 (nM)                                                                     Microonys.                                                                          Prolif.                                                                           IC50 (nM)                                __________________________________________________________________________     ##STR27##                 ++    ++  ++                                        ##STR28##                                                                     ##STR29##                 ++    ++  ++                                        ##STR30##                 ++    ++  ++                                        ##STR31##                 ++    ++  ++                                        ##STR32##                 ++    ++  ++                                        ##STR33##                 ND    +   +                                         ##STR34##                 ++    ++  ++                                        ##STR35##                 ND    ++  ++                                        ##STR36##                 ++    ++  ++                                        ##STR37##                 ++    ++  ++                                        ##STR38##                 ND    ++  ++                                        ##STR39##                 ++    ++  +                                         ##STR40##                 ND    ++  ++                                        ##STR41##                 ++    ++  +-                                        ##STR42##                 +-    +-  +-                                        ##STR43##                 ++    +-  +-                                       __________________________________________________________________________

                  TABLE 8                                                         ______________________________________                                        (SEQ ID NOS 205-222 respectively                                              ______________________________________                                         ##STR44##                                                                     ##STR45##                                                                     ##STR46##                                                                     ##STR47##                                                                     ##STR48##                                                                     ##STR49##                                                                     ##STR50##                                                                     ##STR51##                                                                     ##STR52##                                                                     ##STR53##                                                                     ##STR54##                                                                     ##STR55##                                                                     ##STR56##                                                                     ##STR57##                                                                     ##STR58##                                                                     ##STR59##                                                                    [H]-REGPTLRQWM-[NH.sub.2 ]                                                    [H]-REGPTLRQWLMSRS-[NH.sub.2 ]                                                ______________________________________                                    

                                      TABLE 9                                     __________________________________________________________________________                              EC50 (nM)                                                                     Microonys.                                                                          Prolif.                                                                           IC50 (nM)                                 __________________________________________________________________________    [H]-IEGPTLRQWLAARA-[NH.sub.2 ]  (SEQ ID NO:193)                                                         N.D.  ++  ++                                         ##STR60##                N.D.  ++  ++                                         ##STR61##                ++    ++  ++                                         ##STR62##                ++    ++  ++                                        __________________________________________________________________________

                  TABLE 10                                                        ______________________________________                                         ##STR63##                                                                                         EC50 (nM)                                                Sequence             Cell Prolif.                                                                            IC50 (nM)                                      ______________________________________                                         ##STR64##           ++        ND                                              ##STR65##           ++        ND                                              ##STR66##           ++(++)    ND                                              ##STR67##           ND        ND                                              ##STR68##           ++        ND                                              ##STR69##           ++(++)    ND                                              ##STR70##           ++        ND                                              ##STR71##           ++        ND                                             ______________________________________                                    

EXAMPLE 10

In this example various substitutions were introduced at positions G, P,and W in the cyclized compound (SEQ ID NO:12) ##STR72##

Table 11 lists examples of the substituted compounds that show TPOagonist activity. The substitutions abbreviated in the table are asfollows:

                  TABLE 11                                                        ______________________________________                                        C A D G P T L R E W I S F C - [NH.sub.2 9                                     G              P        W                                                     ______________________________________                                        Sar            Hyp(OBn) Nal                                                   Sar            Hyp(OBn) Nal                                                   Gly            Pro      Trp                                                   Gly            Pro      Trp                                                   Sar            Hyp(OBn) Nal                                                   Gaba           Pro      Trp                                                   Cpr-Gly        Pro      Trp                                                   Sar            Hyp(OBn) Nal                                                   Gly            Pro      Trp                                                   Gly            Pro      Nal                                                   Sar            Pro      Trp                                                   Cpr-Gly        L-Tic    Nal                                                   Gly            D-Tic    D-Trp                                                 Cpr-Gly        D-Tic    Trp                                                   Gaba           Hyp(OBn) Trp                                                   ______________________________________                                    

    __________________________________________________________________________    Proline Replacements                                                          __________________________________________________________________________     ##STR73##                                                                                   ##STR74##                                                                                   ##STR75##                                        L-Pro         D-Pro         L-4-Hyp(OBn)                                       ##STR76##                                                                                   ##STR77##                                                                                   ##STR78##                                        L-Pipecolinic acid                                                                          D-Pipecolinic acid                                                                          L-Azetidine carboxylic                                                        acid                                               ##STR79##                                                                                   ##STR80##                                                                                   ##STR81##                                        L-Tic         D-Tic         L-Oic                                                            ##STR82##                                                                    L-Tiq                                                           __________________________________________________________________________

    __________________________________________________________________________    Tryptophan Replacements                                                       __________________________________________________________________________     ##STR83##                                                                                        ##STR84##                                                                                       ##STR85##                               L-Trp              D-Trp             L-1-Nal                                   ##STR86##                                                                                        ##STR87##                                                                                       ##STR88##                               D-1-Nal            L-2-Nal           D-2-Nal                                   ##STR89##                                                                                        ##STR90##                                                                                       ##STR91##                               L-(Benzothienyl)-alanine                                                                         DL-5-Me-Trp       DL-1-Me-Trp                               ##STR92##                                                                                        ##STR93##                                                                                       ##STR94##                               DL-6-F-Trp         DL-5-F-Trp        DL-5-Br-Trp                                                  ##STR95##                                                                    L-Tic                                                      __________________________________________________________________________

    ______________________________________                                        Glycine Replacements                                                          ______________________________________                                         ##STR96##                                                                                         ##STR97##                                                Glycine             Sarcosine                                                  ##STR98##                                                                                         ##STR99##                                                β-alanine      γ-Aminobutyric acid                                  ##STR100##                                                                                        ##STR101##                                               N-Pentyl glycine    N-Cyclopropyl glycine                                     ______________________________________                                    

EXAMPLE 11

To assess the feasibility of mice as a convenient test species, severalin vitro experiments, designed to measure the activity of the testcompounds on the mouse receptor, have been done. First, marrow cells,harvested from the femurs of 8 to 9 week one Balb/C mice, were incubatedfor 7 days in liquid culture with either rhuTPO or variousconcentrations of the test peptides. At the end of the incubationperiod, the cultures were concentrated by Cytospin, stained foracetylcholinesterase (AChE, a diagnostic of mouse megakaryocytes), andcounted by microscopic analysis. One (1) nM rhuTPO gave rise to theoutgrowth of very large (>40 um) non-adherent cells that stain for AChE.These cells appear to be mature megakaryocytes. From an initial seedingof 10⁶ total marrow cells/ml (in 50 ml cultures) an estimated 1 to 2×10⁶megakaryocytes developed. This responce to TPO was designated as"maximal". Control cultures containing no added growth factors producedvery few AChE-positive cells. Several of the peptide compounds weretested at high concentration in this assay and the results aresummarized in Table 12. Peptide A at 10 uM produced a maximal responseof the mouse marrow. This finding was the first evidence that thispeptide family is active on the murine receptor. In a second experiment,marrow cells were harvested and cultured in semi-solid medium(methylcellulose) containing either no factors, 1 nM rhuTPO, or 10 uMPeptide A. After 7 days in culture, colonies of large cell (presumed tobe megakaryocytes) were counted and grouped into small colonies (3-5cells) or large colonies (greater than 6 cells). The results are shownin Table 13. TPO and the test peptides both produced substantially morecolonies of both sized than did the negative control cultures. Thisindicates that the peptides mimic TPO in their ability to stimulate theexpansion of the Mk precursor cell population.

To obtain a more quantitative comparison of the activity of the testcompounds on murine and human receptors, the muTPO receptor was clonedand transfected into BaF3 cells. A TPO dependent population of cells wasisolated.

                  TABLE 12                                                        ______________________________________                                                        Concentration                                                 Peptide         Tested (nM) Response                                          ______________________________________                                        D               100,000 none                                                  C               40,000  maximal**                                             C ÷ S.A.*   1000    maximal**                                             S.A. alone      1000    none                                                  B               100,000 minimul                                               A               10,000  maximal**                                             TPO (R & D)     1       "maximal"                                             ______________________________________                                         *Streptavidin complexed to biotinylated peptide  concentration of putativ     1:4 complex.                                                                  **Compared to recombinant human TPO                                           **25-30% ACE staining cells on cytopspin                                      No factor cultures  ca. 5% AChE staining cells (lower cellularity)       

                  TABLE 13                                                        ______________________________________                                        Compound             3-5 large cells                                                                          6-12 large cells                              ______________________________________                                        No factors  1        2          1                                             No factors  2        1          1                                             1 nM TPO    #1-1     15         6                                             1 nM TPO    #1-2     12         1                                             1 nM TPO    #2-1     16         8                                             1 nM TPO    #2-2     13         3                                             10 uM Peptide                                                                             #1-1     25         10                                            10 uM Peptide                                                                             #1-2     22         8                                             10 uM Peptide                                                                             #2-1     22         7                                             10 uM Peptide                                                                             #2-2     21         10                                            ______________________________________                                    

The disclosures in this application of all articles and references,including patent documents, are incorporated herein by reference intheir entirety for all purposes.

    __________________________________________________________________________    #             SEQUENCE LISTING                                                - (1) GENERAL INFORMATION:                                                    -    (iii) NUMBER OF SEQUENCES: 232                                           - (2) INFORMATION FOR SEQ ID NO:1:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 5 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                 - Trp Ser Xaa Trp Ser                                                         1               5                                                             - (2) INFORMATION FOR SEQ ID NO:2:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 7 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                 - Xaa Xaa Xaa Xaa Xaa Xaa Xaa                                                 1               5                                                             - (2) INFORMATION FOR SEQ ID NO:3:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 9 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                 - Xaa Gly Xaa Xaa Xaa Xaa Xaa Trp Xaa                                         1               5                                                             - (2) INFORMATION FOR SEQ ID NO:4:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 9 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                 - Xaa Gly Pro Thr Leu Arg Xaa Trp Xaa                                         1               5                                                             - (2) INFORMATION FOR SEQ ID NO:5:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                 - Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Trp Xaa                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:6:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                 - Gly Gly Cys Ala Asp Gly Pro Thr Leu Arg Gl - #u Trp Ile Ser Phe Cys         #                15                                                           - Gly Gly                                                                     - (2) INFORMATION FOR SEQ ID NO:7:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                 - Gly Asn Ala Asp Gly Pro Thr Leu Arg Gln Tr - #p Leu Glu Gly Arg Arg         #                15                                                           - Pro Lys Asn                                                                 - (2) INFORMATION FOR SEQ ID NO:8:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                 - Gly Gly Cys Ala Asp Gly Pro Thr Leu Arg Gl - #u Trp Ile Ser Phe Cys         #                15                                                           - Gly Gly Lys                                                                 - (2) INFORMATION FOR SEQ ID NO:9:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                 - Thr Ile Lys Gly Pro Thr Leu Arg Gln Trp Le - #u Lys Ser Arg Glu His         #                15                                                           - Thr Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:10:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                - Ser Ile Glu Gly Pro Thr Leu Arg Glu Trp Le - #u Thr Ser Arg Thr Pro         #                15                                                           - His Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:11:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                - Leu Ala Ile Glu Gly Pro Thr Leu Arg Gln Tr - #p Leu His Gly Asn Gly         #                15                                                           - Arg Asp Thr                                                                 - (2) INFORMATION FOR SEQ ID NO:12:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                - Cys Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:13:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                - Ile Glu Gly Pro Thr Leu Arg Gln Trp Leu Al - #a Ala Arg Ala                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:14:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 7 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                - Cys Xaa Xaa Xaa Xaa Xaa Xaa                                                 1               5                                                             - (2) INFORMATION FOR SEQ ID NO:15:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                - Gly Gly Cys Thr Leu Arg Glu Trp Leu His Gl - #y Gly Phe Cys Gly Gly         #                15                                                           - (2) INFORMATION FOR SEQ ID NO:16:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 8 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                - Xaa Cys Xaa Xaa Xaa Xaa Xaa Xaa                                             1               5                                                             - (2) INFORMATION FOR SEQ ID NO:17:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 15 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                - Ile Glu Gly Pro Thr Leu Arg Gln Trp Leu Al - #a Ala Arg Ala Lys             #                15                                                           - (2) INFORMATION FOR SEQ ID NO:18:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 15                                                    #/product= "Beta-ala"NFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                - Ile Glu Gly Pro Thr Leu Arg Gln Trp Leu Al - #a Ala Arg Ala Xaa Lys         #                15                                                           - (2) INFORMATION FOR SEQ ID NO:19:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 93 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: cDNA                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..84                                                 -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                - CTC GAG AGC GGG CAG GTG GTG CAT GGG GAG CA - #G GTG GGT GGT GAG GCC           48                                                                          Leu Glu Ser Gly Gln Val Val His Gly Glu Gl - #n Val Gly Gly Glu Ala           #                 15                                                          - TCC GGG GCC GTT AAC GGC CGT GGC CTA GCT GG - #C CAA TAAGTCGAC               #93                                                                           Ser Gly Ala Val Asn Gly Arg Gly Leu Ala Gl - #y Gln                           #             25                                                              - (2) INFORMATION FOR SEQ ID NO:20:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 28 amino                                                          (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                - Leu Glu Ser Gly Gln Val Val His Gly Glu Gl - #n Val Gly Gly Glu Ala         #                 15                                                          - Ser Gly Ala Val Asn Gly Arg Gly Leu Ala Gl - #y Gln                         #             25                                                              - (2) INFORMATION FOR SEQ ID NO:21:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                - Xaa Xaa Xaa Xaa Xaa Thr Leu Arg Glu Trp Le - #u Xaa Xaa Xaa Xaa Xaa         #                15                                                           - Xaa Xaa                                                                     - (2) INFORMATION FOR SEQ ID NO:22:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                - Xaa Xaa Xaa Xaa Xaa Xaa Thr Leu Arg Glu Tr - #p Leu Xaa Xaa Xaa Xaa         #                15                                                           - Xaa Xaa Xaa                                                                 - (2) INFORMATION FOR SEQ ID NO:23:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                - Xaa Xaa Xaa Xaa Xaa Thr Leu Arg Glu Trp Le - #u Xaa Xaa Xaa Xaa Xaa         #                15                                                           - Xaa Xaa                                                                     - (2) INFORMATION FOR SEQ ID NO:24:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                - Xaa Xaa Xaa Xaa Xaa Thr Leu Arg Glu Phe Le - #u Xaa Xaa Xaa Xaa Xaa         #                15                                                           - Xaa Xaa                                                                     - (2) INFORMATION FOR SEQ ID NO:25:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                - Arg Glu Gly Pro Thr Leu Arg Gln Trp Met                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:26:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                - Arg Glu Gly Pro Thr Leu Arg Gln Trp Met                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:27:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                - Ser Arg Gly Met Thr Leu Arg Glu Trp Leu                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:28:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                - Glu Gly Pro Thr Leu Arg Gly Trp Leu Ala                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:29:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                - Arg Glu Gly Gln Thr Leu Lys Glu Trp Leu                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:30:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                - Glu Arg Gly Pro Phe Trp Ala Lys Ala Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:31:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                - Arg Glu Gly Pro Arg Cys Val Met Trp Met                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:32:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                - Cys Ser Gly Leu Thr Leu Arg Glu Trp Leu Va - #l Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:33:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                - Cys Leu Thr Gly Pro Phe Val Thr Gln Trp Le - #u Tyr Glu Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:34:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                - Cys Gly Glu Gly Leu Thr Leu Thr Gln Trp Le - #u Glu His Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:35:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                - Cys Arg Ala Gly Pro Thr Leu Leu Glu Trp Le - #u Thr Leu Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:36:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                - Cys Arg Ala Gly Pro Thr Leu Leu Glu Trp Le - #u Thr Leu Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:37:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                - Cys Arg Gln Gly Pro Thr Leu Thr Ala Trp Le - #u Leu Glu Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:38:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                - Cys Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:39:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                - Cys Glu Leu Val Gly Pro Ser Leu Met Ser Tr - #p Leu Thr Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:40:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                - Cys Gly Thr Glu Gly Pro Thr Leu Ser Thr Tr - #p Leu Asp Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:41:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                - Cys Asp Gln Leu Gly Val Thr Leu Ser Arg Tr - #p Leu Glu Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:42:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                - Ser Gly Thr Gly Leu Thr Leu Arg Glu Trp Le - #u Gly Ser Phe Ser Leu         #                15                                                           - Leu Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:43:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                - Cys Pro Glu Gly Pro Thr Leu Leu Gln Trp Le - #u Lys Arg Gly Tyr Ser         #                15                                                           - Ser Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:44:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                                - Arg Gly Asp Gly Pro Thr Leu Ser Gln Trp Le - #u Tyr Ser Leu Met Ile         #                15                                                           - Met Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:45:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                                - Met Val Ala Gly Pro Thr Leu Arg Glu Phe Il - #e Ala Ser Leu Pro Ile         #                15                                                           - His Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:46:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                                - Ser Met Gln Gly Pro Thr Phe Arg Glu Trp Va - #l Ser Met Met Lys Val         #                15                                                           - Leu Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:47:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                                - Ser Val Gln Cys Gly Pro Thr Leu Arg Gln Tr - #p Leu Ala Ala Arg Asn         #                15                                                           - His Leu Ser                                                                 - (2) INFORMATION FOR SEQ ID NO:48:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                                - Gly Asn Ala Asp Gly Pro Thr Leu Arg Gln Tr - #p Leu Glu Gly Arg Arg         #                15                                                           - Pro Lys Asn                                                                 - (2) INFORMATION FOR SEQ ID NO:49:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                                - Ser Val Arg Cys Gly Pro Thr Leu Arg Gln Tr - #p Leu Ala Ala Arg Thr         #                15                                                           - His Leu Ser                                                                 - (2) INFORMATION FOR SEQ ID NO:50:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                                - Leu Ala Ile Glu Gly Pro Thr Leu Arg Gln Tr - #p Leu His Gly Asn Gly         #                15                                                           - Arg Asp Thr                                                                 - (2) INFORMATION FOR SEQ ID NO:51:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                                - His Gly Arg Val Gly Pro Thr Leu Arg Glu Tr - #p Lys Thr Gln Val Ala         #                15                                                           - Thr Lys Lys                                                                 - (2) INFORMATION FOR SEQ ID NO:52:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                                - Cys Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:53:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                                - Ile Ser Asp Gly Pro Thr Leu Lys Glu Trp Le - #u Ser Val Thr Arg Gly         #                15                                                           - Ala Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:54:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                                - Ser Ile Glu Gly Pro Thr Leu Arg Glu Trp Le - #u Thr Ser Arg Thr Pro         #                15                                                           - His Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:55:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                                - Thr Ile Lys Gly Pro Thr Leu Arg Gln Trp Le - #u Lys Ser Arg Glu His         #                15                                                           - Thr Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:56:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                                - Gly Asn Ala Asp Gly Pro Thr Leu Arg Gln Tr - #p Leu Glu Gly Arg Arg         #                15                                                           - Pro Lys Asn                                                                 - (2) INFORMATION FOR SEQ ID NO:57:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                                - Ser Ile Glu Gly Pro Thr Leu Arg Glu Trp Le - #u Thr Ser Arg Thr Pro         #                15                                                           - His Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:58:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                                - Ile Ser Asp Gly Pro Thr Leu Lys Glu Trp Le - #u Ser Val Thr Arg Gly         #                15                                                           - Ala Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:59:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                                - Cys Ser Leu Glu Asp Leu Arg Lys Arg Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:60:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                                - Cys Arg Arg Ser Glu Leu Leu Glu Arg Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:61:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                                - Cys Thr Phe Lys Gln Phe Leu Asp Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:62:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                                - Cys Thr Arg Gly Glu Trp Leu Arg Cys Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:63:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                                - Cys Thr Leu Arg Gln Trp Leu Gln Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:64:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                                - Cys Thr Leu Glu Glu Leu Arg Ala Cys Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:65:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                                - Cys Thr Arg Glu Glu Leu Met Arg Leu Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:66:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                                - Cys Gln Arg Ala Asp Leu Ile Asn Phe Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:67:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                                - Cys Asn Arg Asn Asp Leu Leu Leu Phe Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:68:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                                - Cys Thr Arg Thr Glu Trp Leu His Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:69:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 9 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                                - Cys Thr Leu Glu Phe Met Asn Gly Cys                                         1               5                                                             - (2) INFORMATION FOR SEQ ID NO:70:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                                - Cys Ser Leu Gly Glu Leu Arg Arg Leu Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:71:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:                                - Cys Asn Ile Asn Gln Leu Arg Ser Ile Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:72:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:                                - Cys Thr Met Arg Gln Phe Leu Val Cys Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:73:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:                                - Cys Thr Arg Ser Glu Trp Leu Glu Arg Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:74:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:                                - Cys Thr Leu His Glu Tyr Leu Ser Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:75:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:                                - Cys Thr Arg Glu Glu Leu Leu Arg Gln Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:76:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:                                - Cys Thr Phe Arg Glu Phe Val Asn Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:77:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:                                - Cys Ser Arg Ala Asp Phe Leu Ala Ala Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:78:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78:                                - Cys Ser Cys Ala Gln Val Val Gln Cys Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:79:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:                                - Cys Thr Leu Arg Gln Trp Ile Leu Leu Gly Me - #t Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:80:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:                                - Cys Thr Leu Arg Glu Trp Leu His Gly Gly Ph - #e Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:81:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:                                - Cys Thr Leu Arg Ala Trp Leu Met Ser Glu Th - #r Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:82:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:                                - Cys Thr Leu Arg Ala Trp Leu Met Glu Ser Cy - #s Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:83:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:                                - Cys Thr Phe Gln Val Trp Lys Leu Ala Arg As - #n Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:84:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84:                                - Cys Leu Leu Arg Glu Trp Leu Asp Xaa Arg Th - #r Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:85:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85:                                - Cys Val Leu Arg Glu Trp Leu Leu Xaa Xaa Se - #r Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:86:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86:                                - Cys Leu Leu Ser Glu Phe Leu Ala Gly Gln Gl - #n Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:87:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:                                - Cys Ser Leu Arg Gln Tyr Leu Asp Phe Gly Le - #u Gly Ser Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:88:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88:                                - Cys Thr Leu Gln Glu Leu Lys Gln Ser Ser Le - #u Tyr Glu Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:89:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89:                                - Cys Asp Leu Ser Glu Leu Lys Thr His Gly Ty - #r Ala Tyr Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:90:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:90:                                - Cys Lys Leu Ser Asp Trp Leu Met Asn Gly Va - #l Ala Ala Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:91:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:91:                                - Cys Ser Leu Gln Glu Phe Leu Ser His Gly Gl - #y Tyr Val Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:92:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:92:                                - Cys Ser Leu Lys Glu Phe Leu His Ser Gly Le - #u Met Gln Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:93:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:93:                                - Cys Thr Phe Arg Gln Leu Leu Glu Tyr Gly Va - #l Ser Ser Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:94:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94:                                - Cys Thr Met Arg Glu Phe Leu Val Ala Ser Gl - #y Val Ala Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:95:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:95:                                - Cys Thr Leu Ala Glu Phe Leu Ala Ser Gly Va - #l Glu Gln Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:96:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96:                                - Cys Thr Leu Ala Glu Phe Leu Ala Ser Gly Va - #l Glu Gln Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:97:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:97:                                - Cys Thr Leu Lys Glu Trp Leu Val Ser His Gl - #u Val Trp Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:98:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98:                                - Cys Thr Leu Arg Glu Phe Leu Ser Leu Gly Me - #t Asn Ala Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:99:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99:                                - Cys Thr Leu Arg Glu Phe Leu Asp Pro Thr Th - #r Ala Val Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:100:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100:                               - Cys Ser Leu Leu Glu Phe Leu Ala Leu Gly Va - #l Ala Leu Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:101:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:                               - Gly Gly Gly Arg Gly Cys Thr Leu Lys Gln Tr - #p Lys Gln Gly Asp Cys         #                15                                                           - Gly Arg Ser                                                                 - (2) INFORMATION FOR SEQ ID NO:102:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:                               - Cys Asn Arg Ser Gln Leu Leu Ala Ala Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:103:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103:                               - Cys Thr Leu Gln Gln Trp Leu Ser Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:104:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104:                               - Cys Thr Leu Arg Glu Phe Lys Ala Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:105:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:105:                               - Cys Thr Arg Ala Gln Phe Leu Lys Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:106:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106:                               - Cys Thr Leu Arg Glu Phe Asn Arg Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:107:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107:                               - Cys Thr Leu Ser Asp Phe Lys Arg Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:108:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:108:                               - Cys Thr Phe Arg Gln Trp Lys Glu Ala Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:109:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:109:                               - Cys Thr Leu Ser Glu Phe Arg Gly Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:110:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110:                               - Cys Thr Leu Gln Glu Phe Leu Glu Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:111:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111:                               - Cys Thr Leu Gln Gln Trp Lys Asp Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:112:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:112:                               - Cys Thr Arg Ser Gln Trp Leu Glu Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:113:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:113:                               - Cys Ser Leu Gln Glu Phe Lys His Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:114:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114:                               - Cys Thr Leu Gly Glu Trp Lys Arg Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:115:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 11 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:115:                               - Cys Thr Leu Trp Gly Cys Gly Lys Arg Gly Cy - #s                             #                10                                                           - (2) INFORMATION FOR SEQ ID NO:116:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:116:                               - Cys Thr Leu Gln Glu Trp Arg Gly Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:117:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:117:                               - Cys Thr Arg Leu Ser Gly Cys Trp Leu Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:118:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:118:                               - Cys Thr Arg Thr Gln Trp Leu Leu Asp Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:119:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:119:                               - Cys Thr Leu Ala Glu Phe Arg Arg Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:120:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:120:                               - Cys Thr Ser Thr Gln Trp Leu Leu Ala Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:121:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121:                               - Cys Ser Arg Ser Gln Phe Leu Arg Ser Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:122:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122:                               - Cys Thr Leu Arg Glu Trp Leu Glu Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:123:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:123:                               - Cys Thr Leu Arg Glu Phe Leu Leu Met Gly Al - #a Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:124:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:124:                               - Cys Thr Leu Lys Glu Trp Leu Leu Trp Ser Se - #r Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:125:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125:                               - Cys Thr Leu Leu Glu Trp Leu Arg Asn Pro Va - #l Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:126:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:126:                               - Cys Thr Leu Arg Gln Trp Leu Gly Asp Ala Tr - #p Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:127:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:127:                               - Cys Thr Leu Gly Gln Trp Leu Gln Met Gly Me - #t Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:128:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:128:                               - Cys Thr Leu Arg Glu Trp Val Phe Ala Gly Le - #u Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:129:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:129:                               - Cys Leu Leu Leu Glu Phe Leu Ser Gly Ala As - #p Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:130:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:130:                               - Cys Thr Leu Gly Glu Phe Leu Ala Gly His Le - #u Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:131:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131:                               - Cys Arg Leu Arg Glu Phe Leu Val Asp Leu Th - #r Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:132:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:132:                               - Cys Ser Phe Arg Ser Trp Leu Val Asp Gln Th - #r Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:133:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133:                               - Cys Thr Leu Arg Glu Trp Leu Glu Asp Leu Gl - #y Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:134:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:134:                               - Cys Thr Leu Gln Asp Trp Leu Val Ser Trp Th - #r Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:135:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:135:                               - Cys Thr Leu Ser Glu Trp Leu Ser Glu Leu Se - #r Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:136:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:136:                               - Cys Thr Leu Met Gln Trp Leu Gly Gly Trp Pr - #o Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:137:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:137:                               - Cys Thr Leu Arg Glu Trp Leu Ser Tyr Gly Th - #r Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:138:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:138:                               - Cys Thr Leu Gln Glu Trp Leu Ser Gly Gly Le - #u Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:139:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:139:                               - Gly Ser His Gly Cys Thr Leu Arg Glu Trp Le - #u Cys Met Lys Ile Val         #                15                                                           - Pro Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:140:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:140:                               - Gln Trp Gln Gly Cys Thr Leu Arg Asp Cys Il - #e Leu Arg Gly Val Phe         #                15                                                           - Trp Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:141:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:141:                               - Ser Val Asn Ser Cys Thr Leu Arg Glu Phe Le - #u Thr Gly Cys Arg Val         #                15                                                           - Phe Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:142:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:142:                               - Ser Tyr Asp Gly Cys Thr Leu Arg His Trp Le - #u Met Asp Ile Tyr Gly         #                15                                                           - Asp Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:143:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:143:                               - Gln Arg Ser Gly Cys Thr Leu Arg Asp Trp Va - #l Leu Leu Asn Cys Leu         #                15                                                           - Ala Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:144:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:144:                               - Asn Tyr Arg Gly Cys Thr Leu Ser Gln Trp Va - #l Ser Glu Gln Ile Val         #                15                                                           - Gly Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:145:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145:                               - Gly Arg Ser Gly Cys Thr Leu Arg Glu Tyr Le - #u Gly Gly Met Cys Tyr         #                15                                                           - Leu Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:146:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146:                               - Ala Ser Trp Tyr Cys Thr Val Pro Glu Leu Me - #t Glu Met Gln Leu Pro         #                15                                                           - Glu Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:147:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:147:                               - Gly Ser Thr Gly Cys Thr Leu Arg Glu Xaa Le - #u His Met Leu Gly Leu         #                15                                                           - Asp Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:148:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:148:                               - Ala Cys Glu Gly Cys Thr Leu Arg Gln Trp Le - #u Glu Tyr Val Arg Val         #                15                                                           - Gly Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:149:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:149:                               - Ala Gln Arg Gly Cys Thr Leu Gln Tyr Phe Va - #l Ser Tyr Gly Xaa Asp         #                15                                                           - Met Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:150:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:150:                               - Gly Val Cys Gly Cys Thr Leu Arg Glu Phe Le - #u Ala Ile Pro His Thr         #                15                                                           - Ser Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:151:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:151:                               - Ser Glu Gly Gly Cys Thr Leu Arg Glu Trp Va - #l Ala Ser Ser Leu Ala         #                15                                                           - Asn Cys                                                                     - (2) INFORMATION FOR SEQ ID NO:152:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:152:                               - Ser Asn Ser Arg Cys Thr Leu Arg Glu Trp Il - #e Ile Gln Gly Cys Asp         #                15                                                           - Phe Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:153:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:153:                               - Ser Asn Ser Arg Cys Thr Leu Arg Glu Trp Il - #e Ile Gln Gly Cys Asp         #                15                                                           - Phe Ser                                                                     - (2) INFORMATION FOR SEQ ID NO:154:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:154:                               - Cys Leu Gly Cys Thr Leu Ser Gln Trp Arg Ly - #s Arg Thr Arg Cys Asp         #                15                                                           - Thr His                                                                     - (2) INFORMATION FOR SEQ ID NO:155:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 17 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:155:                               - Tyr Arg Gly Cys Ser Arg Ala Gln Leu Leu Gl - #y Gly Glu Cys Arg Lys         #                15                                                           - Lys                                                                         - (2) INFORMATION FOR SEQ ID NO:156:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 17 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:156:                               - Gly Arg Gly Cys Thr Leu Lys Gln Trp Lys Gl - #n Gly Asp Cys Gly Arg         #                15                                                           - Ser                                                                         - (2) INFORMATION FOR SEQ ID NO:157:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:157:                               - Val Arg Gly Gly Cys Ala Leu Arg Asp Trp Va - #l Ala Gly Glu Cys Phe         #                15                                                           - Asp Trp Thr                                                                 - (2) INFORMATION FOR SEQ ID NO:158:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:158:                               - Leu Trp Arg Gly Cys Thr Leu Asn Gly Phe Ly - #s Ser Arg His Cys Gly         #                15                                                           - Ser Pro Glu                                                                 - (2) INFORMATION FOR SEQ ID NO:159:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 13 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:159:                               - Cys Thr Leu Arg Ser Trp Lys His Arg Gly Cy - #s Ala Pro                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:160:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 17 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:160:                               - Gly Arg Gly Cys Thr Arg Ala Gln Trp Leu Al - #a Gly Cys Cys Thr Gly         #                15                                                           - His                                                                         - (2) INFORMATION FOR SEQ ID NO:161:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:161:                               - Arg Ala Gly Cys Thr Leu Arg Glu Phe Arg Ly - #s Gly Cys Leu Ala Leu         #                15                                                           - (2) INFORMATION FOR SEQ ID NO:162:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 17 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:162:                               - Lys Arg Gly Cys Thr Leu Ala Glu Met Ile Ar - #g Gly Cys Asn Arg Ser         #                15                                                           - Asn                                                                         - (2) INFORMATION FOR SEQ ID NO:163:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 17 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:163:                               - Gly Arg Gly Cys Thr Leu Lys Gln Trp Lys Gl - #n Gly Asp Cys Gly Arg         #                15                                                           - Ser                                                                         - (2) INFORMATION FOR SEQ ID NO:164:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:164:                               - Arg Trp Arg Gly Cys Ser Leu Ala Lys Leu Ly - #s Lys Gly Ala Ala Cys         #                15                                                           - Gly Arg Gly                                                                 - (2) INFORMATION FOR SEQ ID NO:165:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:165:                               - Arg Gly Gly Cys Thr Leu Arg Glu Trp Arg Ar - #g Val Arg Val Ile Asn         #                15                                                           - (2) INFORMATION FOR SEQ ID NO:166:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 17 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:166:                               - Gly Arg Gly Cys Thr Leu Lys Gln Trp Lys Gl - #n Gly Asp Cys Gly Arg         #                15                                                           - Ser                                                                         - (2) INFORMATION FOR SEQ ID NO:167:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 17 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:167:                               - Arg Tyr Gly Cys Thr Arg His Gln Trp Leu Va - #l Gly Thr Cys Val Arg         #                15                                                           - His                                                                         - (2) INFORMATION FOR SEQ ID NO:168:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 8 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:168:                               - Gly Pro Thr Leu Arg Gln Trp Leu                                             1               5                                                             - (2) INFORMATION FOR SEQ ID NO:169:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 11 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: other nucleic acid                                  #= "oligonucleotide"PTION: /desc                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:169:                               #       11                                                                    - (2) INFORMATION FOR SEQ ID NO:170:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 11 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: other nucleic acid                                  #= "oligonucleotide"PTION: /desc                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:170:                               #       11                                                                    - (2) INFORMATION FOR SEQ ID NO:171:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 25 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: other nucleic acid                                  #= "oligonucleotide"PTION: /desc                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:171:                               #               25 AAGT AAAGC                                                 - (2) INFORMATION FOR SEQ ID NO:172:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 1                                                     #/product= "Pen"HER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:172:                               - Xaa Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:173:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 13 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:173:                               - Ala Asp Gly Pro Thr Leu Arg Glu Trp Ile Se - #r Phe Cys                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:174:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 1                                                     #/product= "Homocys"INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:174:                               - Xaa Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:175:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 1                                                     #/product= "D-Cys"R INFORMATION:                                              -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 14                                                    #/product= "D-Cys"R INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:175:                               - Xaa Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Xaa                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:176:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 14                                                    #/product= "D-Cys"R INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:176:                               - Cys Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Xaa                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:177:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 1                                                     #/product= "D-Pen"R INFORMATION:                                              -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 14                                                    #/product= "D-Cys"R INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:177:                               - Xaa Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Xaa                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:178:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 1                                                     #/product= "Homocys"INFORMATION:                                              -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 14                                                    #/product= "Homocys"INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:178:                               - Xaa Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Xaa                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:179:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 13 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 13                                                    #/product= "Homocys"INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:179:                               - Ala Asp Gly Pro Thr Leu Arg Glu Trp Ile Se - #r Phe Xaa                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:180:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 13 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 13                                                    #/product= "Pen"HER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:180:                               - Ala Asp Gly Pro Thr Leu Arg Glu Trp Ile Se - #r Phe Xaa                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:181:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:181:                               - Lys Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Glu                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:182:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:182:                               - Glu Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Lys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:183:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 13 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:183:                               - Ala Asp Gly Pro Thr Leu Arg Glu Trp Ile Se - #r Phe Glu                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:184:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 1                                                     #/product= "Pen"HER INFORMATION:                                              -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 14                                                    #/product= "Pen"HER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:184:                               - Xaa Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe Xaa                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:185:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 15 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:185:                               - Cys Ile Glu Gly Pro Thr Leu Arg Gln Trp Le - #u Ala Ala Arg Ala             #                15                                                           - (2) INFORMATION FOR SEQ ID NO:186:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 13 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:186:                               - Cys Ala Asp Gly Pro Thr Leu Arg Glu Trp Il - #e Ser Phe                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:187:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:187:                               - Ser Val Gln Cys Gly Pro Thr Leu Arg Gln Tr - #p Leu Ala Ala Arg Asn         #                15                                                           - His Leu Ser                                                                 - (2) INFORMATION FOR SEQ ID NO:188:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:188:                               - Met Val Gly Pro Thr Leu Arg Ser Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:189:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:189:                               - Asp Gly Pro Thr Leu Arg Glu Trp Ile Ser Ph - #e Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:190:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 11 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:190:                               - Gly Pro Thr Leu Arg Glu Trp Ile Ser Phe Cy - #s                             #                10                                                           - (2) INFORMATION FOR SEQ ID NO:191:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:191:                               - Pro Thr Leu Arg Glu Trp Ile Ser Phe Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:192:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 9 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:192:                               - Thr Leu Arg Glu Trp Ile Ser Phe Cys                                         1               5                                                             - (2) INFORMATION FOR SEQ ID NO:193:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:193:                               - Ile Glu Gly Pro Thr Leu Arg Gln Trp Leu Al - #a Ala Arg Ala                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:194:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:194:                               - Cys Ile Glu Gly Pro Thr Leu Arg Gln Trp Le - #u Ala Ala Arg Ala Cys         #                15                                                           - (2) INFORMATION FOR SEQ ID NO:195:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:195:                               - Ile Glu Gly Pro Thr Leu Arg Gln Trp Leu Al - #a Ala Arg Lys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:196:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 15 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 14                                                    #/product= "Beta-ala"NFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:196:                               - Ile Glu Gly Pro Thr Leu Arg Gln Trp Leu Al - #a Ala Arg Xaa Lys             #                15                                                           - (2) INFORMATION FOR SEQ ID NO:197:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 13 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:197:                               - Ile Glu Gly Pro Thr Leu Arg Gln Trp Leu Al - #a Ala Lys                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:198:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 13                                                    #/product= "Beta-ala"NFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:198:                               - Ile Glu Gly Pro Thr Leu Arg Gln Trp Leu Al - #a Ala Xaa Lys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:199:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:199:                               - Glu Gly Pro Thr Leu Arg Gln Trp Leu Ala Al - #a Arg Ala Lys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:200:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 15 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 14                                                    #/product= "Beta-ala"NFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:200:                               - Glu Gly Pro Thr Leu Arg Gln Trp Leu Ala Al - #a Arg Ala Xaa Lys             #                15                                                           - (2) INFORMATION FOR SEQ ID NO:201:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 13 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:201:                               - Glu Gly Pro Thr Leu Arg Gln Trp Leu Ala Al - #a Arg Lys                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:202:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 13                                                    #/product= "Beta-ala"NFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:202:                               - Glu Gly Pro Thr Leu Arg Gln Trp Leu Ala Al - #a Arg Xaa Lys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:203:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:203:                               - Glu Gly Pro Thr Leu Arg Gln Trp Leu Ala Al - #a Lys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:204:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 13 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 12                                                    #/product= "Beta-ala"NFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:204:                               - Glu Gly Pro Thr Leu Arg Gln Trp Leu Ala Al - #a Xaa Lys                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:205:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:205:                               - Cys Thr Arg Ala Gln Phe Leu Lys Gly Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:206:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:206:                               - Cys Asn Ile Asn Gln Leu Arg Ser Ile Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:207:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:207:                               - Cys Asn Arg Ser Gln Leu Leu Ala Ala Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:208:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:208:                               - Cys Thr Ser Thr Gln Trp Leu Leu Ala Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:209:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:209:                               - Cys Gln Arg Ala Asp Leu Ile Asn Phe Cys                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:210:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:210:                               - Cys Leu Leu Ser Glu Phe Leu Ala Gly Gln Gl - #n Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:211:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:211:                               - Cys Thr Phe Gln Val Trp Lys Leu Ala Arg As - #n Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:212:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 12 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:212:                               - Cys Thr Leu Gly Gln Trp Leu Gln Met Gly Me - #t Cys                         #                10                                                           - (2) INFORMATION FOR SEQ ID NO:213:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:213:                               - Cys Leu Thr Gly Pro Phe Val Thr Gln Trp Le - #u Tyr Glu Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:214:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:214:                               - Cys Thr Leu Arg Glu Phe Leu Asp Pro Thr Th - #r Ala Val Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:215:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:215:                               - Cys Gly Thr Glu Gly Pro Thr Leu Ser Thr Tr - #p Leu Asp Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:216:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:216:                               - Cys Glu Leu Val Gly Pro Ser Leu Met Ser Tr - #p Leu Thr Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:217:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:217:                               - Cys Ser Leu Lys Glu Phe Leu His Ser Gly Le - #u Met Gln Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:218:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:218:                               - Cys Thr Leu Ala Glu Phe Leu Ala Ser Gly Va - #l Glu Gln Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:219:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             #ID NO:219:xi) SEQUENCE DESCRIPTION: SEQ                                      - Cys Thr Leu Lys Glu Trp Leu Val Ser His Gl - #u Val Trp Cys                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:220:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:220:                               - Cys Ile Glu Gly Pro Thr Leu Arg Gln Trp Le - #u Ala Ala Arg Ala Cys         #                15                                                           - (2) INFORMATION FOR SEQ ID NO:221:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 10 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:221:                               - Arg Glu Gly Pro Thr Leu Arg Gln Trp Met                                     #                10                                                           - (2) INFORMATION FOR SEQ ID NO:222:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:222:                               - Arg Glu Gly Pro Thr Leu Arg Gln Trp Leu Me - #t Ser Arg Ser                 #                10                                                           - (2) INFORMATION FOR SEQ ID NO:223:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 92 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: cDNA                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..63                                                 -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:223:                               - CTC GAG AGC GGG CAG GTG GTG CAT GGG GAG CA - #G GTG GGT GGT GAG GCC           48                                                                          Leu Glu Ser Gly Gln Val Val His Gly Glu Gl - #n Val Gly Gly Glu Ala           #                 15                                                          - TCC GGA GGT GGT NNK TAACTAAGTA AAGCTGGCCA ATAAGTCGA - #                     # 92                                                                          Ser Gly Gly Gly Xaa                                                                        20                                                               - (2) INFORMATION FOR SEQ ID NO:224:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 21 amino                                                          (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:224:                               - Leu Glu Ser Gly Gln Val Val His Gly Glu Gl - #n Val Gly Gly Glu Ala         #                 15                                                          - Ser Gly Gly Gly Xaa                                                                      20                                                               - (2) INFORMATION FOR SEQ ID NO:225:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 122 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: cDNA                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..120                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:225:                               - CAG ACT AAT TCG AGC TCG AAC AAC AAC AAC AA - #T AAC AAT AAC AAC AAC           48                                                                          Gln Thr Asn Ser Ser Ser Asn Asn Asn Asn As - #n Asn Asn Asn Asn Asn           #                 15                                                          - CTC GGG ATC GAG GGA AGG ACC GGT CAC GTG GC - #C CGG GAA TTC GGA TCC           96                                                                          Leu Gly Ile Glu Gly Arg Thr Gly His Val Al - #a Arg Glu Phe Gly Ser           #             30                                                              #             122  TG CAG GCA AGC TT                                          Ser Arg Val Asp Leu Gln Ala Ser                                               #         40                                                                  - (2) INFORMATION FOR SEQ ID NO:226:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 40 amino                                                          (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:226:                               - Gln Thr Asn Ser Ser Ser Asn Asn Asn Asn As - #n Asn Asn Asn Asn Asn         #                 15                                                          - Leu Gly Ile Glu Gly Arg Thr Gly His Val Al - #a Arg Glu Phe Gly Ser         #             30                                                              - Ser Arg Val Asp Leu Gln Ala Ser                                             #         40                                                                  - (2) INFORMATION FOR SEQ ID NO:227:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 51 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: cDNA                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..21                                                 -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:227:                               - GGA AGG ACC GGA GGT GGT NNK TAACTAAGTA AAGCTGGCC - #A ATAAGTCGAC              51                                                                          Gly Arg Thr Gly Gly Gly Xaa                                                     1               5                                                           - (2) INFORMATION FOR SEQ ID NO:228:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 7 amino                                                           (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:228:                               - Gly Arg Thr Gly Gly Gly Xaa                                                   1               5                                                           - (2) INFORMATION FOR SEQ ID NO:229:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 93 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: cDNA                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..84                                                 -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:229:                               - GAA GCG GCG ATG GCG GAG CTG AAT TAC ATT CC - #C CGG TCG CAG GAG GCC           48                                                                          Glu Ala Ala Met Ala Glu Leu Asn Tyr Ile Pr - #o Arg Ser Gln Glu Ala           #                 15                                                          - TCC GGG GCC GTT AAC GGC CGT GGC CTA GCT GG - #C CAA TAAGTCGAC               #93                                                                           Ser Gly Ala Val Asn Gly Arg Gly Leu Ala Gl - #y Gln                           #             25                                                              - (2) INFORMATION FOR SEQ ID NO:230:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 28 amino                                                          (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:230:                               - Glu Ala Ala Met Ala Glu Leu Asn Tyr Ile Pr - #o Arg Ser Gln Glu Ala         #                 15                                                          - Ser Gly Ala Val Asn Gly Arg Gly Leu Ala Gl - #y Gln                         #             25                                                              - (2) INFORMATION FOR SEQ ID NO:231:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 126 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: cDNA                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..96                                                 -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:231:                               - GAA GCG GCG ATG GCG GAG CTG AAT TAC ATT CC - #C CGG TCG CAG GAG GCC           48                                                                          Glu Ala Ala Met Ala Glu Leu Asn Tyr Ile Pr - #o Arg Ser Gln Glu Ala           #                 15                                                          - TCC GGA GGT GGT NNK NNK NNK NNK NNK NNK NN - #K NNK NNK NNK NNK NNK           96                                                                          Ser Gly Gly Gly Xaa Xaa Xaa Xaa Xaa Xaa Xa - #a Xaa Xaa Xaa Xaa Xaa           #             30                                                              #          126     GCCA ATAAGTCGAC                                            - (2) INFORMATION FOR SEQ ID NO:232:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 32 amino                                                          (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:232:                               - Glu Ala Ala Met Ala Glu Leu Asn Tyr Ile Pr - #o Arg Ser Gln Glu Ala         #                 15                                                          - Ser Gly Gly Gly Xaa Xaa Xaa Xaa Xaa Xaa Xa - #a Xaa Xaa Xaa Xaa Xaa         #             30                                                              __________________________________________________________________________

What is claimed is:
 1. A compound that binds to thrombopoietin receptor, said compound having:(1) a molecular weight of less than about 8000 daltons, and (2) a binding affinity to thrombopoietin receptor as expressed by an IC₅₀ of no more than about 100 μm.
 2. The compound of claim 1, wherein said compound is a peptide, and,wherein from zero to all of the --C(O)NH-- linkages of the peptide have been replaced by a linkage selected from the group consisting of a --CH₂ OC(O)NR-- linkage; a phosphonate linkage; a --CH₂ S(O)₂ NR-- linkage; a --CH₂ NR-- linkage; a --C(O)NR⁶ -- linkage; and a --NHC(O)NH-- linkage; and wherein R is hydrogen or lower alkyl and R⁶ is lower alkyl, further wherein the N-terminus of said peptide is selected from the group consisting of a --NRR¹ group; a --NRC(O)R group; a --NRC(O)OR group; a --NRS(O)₂ R group; a --NHC(O)NHR group; a succinimide group; a benzyloxycarbonyl-NH-- group; and a benzyloxycarbonyl-NH-- group having from 1 to 3 substituents on the phenyl ring selected from the group consisting of lower alkyl, lower alkoxy, chloro, and bromo; and wherein R and R¹ are independently selected from the group consisting of hydrogen and lower alkyl, and still further wherein the C-terminus of said peptide has the formula --C(O)R² where R² is selected from the group consisting of hydroxy, lower alkoxy, and --NR³ R⁴ where R³ and R⁴ are independently selected from the group consisting of hydrogen and lower alkyl and where the nitrogen atom of the --NR³ R⁴ group can optionally be the amine group of the N-terminus of the peptide so as to form a cyclic peptide, and physiologically acceptable salts thereof.
 3. A pharmaceutical composition comprising the compound of claim 1 in combination with a pharmaceutically acceptable carrier.
 4. A method for treating a patient suffering from a disorder that is susceptible to treatment with a thrombopoietin agonist, comprising admistering to the patient, a therapeutically effective dose or amount of a compound of claim
 1. 5. The method of claim 4, wherein the compound administered to the patient is a peptide, and,wherein from zero to all of the --C(O)NH-- linkages of the peptide have been replaced by a linkage selected from the group consisting of a --CH₂ OC(O)NR-- linkage; a phosphonate linkage; a --CH₂ S(O)₂ NR-- linkage; a --CH₂ NR-- linkage; a --C(O)NR⁶ -- linkage; and a --NHC(O)NH-- linkage; and wherein R is hydrogen or lower alkyl and R⁶ is lower alkyl, further wherein the N-terminus of said peptide is selected from the group consisting of a --NRR¹ group; a --NRC(O)R group; a --NRC(O)OR group; a --NRS(O)₂ R group; a --NHC(O)NHR group; a succinimide group; a benzyloxycarbonyl-NH-- group; and a benzyloxycarbonyl-NH-- group having from 1 to 3 substituents on the phenyl ring selected from the group consisting of lower alkyl, lower alkoxy, chloro, and bromo; and wherein R and R¹ are independently selected from the group consisting of hydrogen and lower alkyl, and still further wherein the C-terminus of said peptide has the formula --C(O)R² where R² is selected from the group consisting of hydroxy, lower alkoxy, and --NR³ R⁴ where R³ and R⁴ are independently selected from the group consisting of hydrogen and lower alkyl and where the nitrogen atom of the --NR³ R⁴ group can optionally be the amine group of the N-terminus of the peptide so as to form a cyclic peptide, and physiologically acceptable salts thereof.
 6. The compound of claim 2, wherein said compound comprises a sequence of amino acids (SEQ ID NO:2):

    X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7

where X₁ is C, L, M, P, Q, V; X₂ is F, K, L, N, Q, R, S, T or V; X₃ is C, F, I, L, M, R, S, V or W; X₄ is any of the 20 genetically coded L-amino acids; X₅ is A, D, E, G, K, M, Q, R, S, T, V or Y; X₆ is C, F, G, L, M, S, V, W or Y; and X₇ is C, G, I, K, L, M, N, R or V.
 7. The compound of claim 6, wherein said sequence of amino acids is cyclized.
 8. The compound of claim 6, wherein said sequence of amino acids is dimerized.
 9. The compound of claim 6, wherein the compound comprises the sequence of amino acids (SEQ ID NO:14)

    C X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7

where X₂ is K, L, N, Q, R, S, T or V; X₃ is C, F, I, L, M, R, S or V; X₄ is any of the 20 genetically coded L-amino acids; X₅ is A, D, E, G, S, V or Y; X₆ is C, F, G, L, M, S, V, W or Y; and X₇ is C, G, I, K, L, M, N, R or V.
 10. The compound of claim 8, wherein X₄ is A, E, G, H, K, L, M, P, Q, R, S, T, or W.
 11. The compound of claim 10, wherein X₂ is S or T; X₃ is L or R; X₄ is R; X₅ is D, E, or G; X₆ is F, L, or W; and X₇ is I, K, L, R, or V.
 12. The compound of claim 9, wherein said compound comprises a sequence of amino acids (SEQ ID NO:16):

    X.sub.8 C X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7

where X₂ is F, K, L, N, Q, R, S, T or V; X₃ is C, F, I, L, M, R, S, V or W; X₄ is any of the 20 genetically coded L-amino acids; X₅ is A, D, E, G, K, M, Q, R, S, T, V or Y; X₆ is C, F, G, L, M, S, V, W or Y; X₇ is C, G, I, K, L, M, N, R or V; and X₈ is any of the 20 genetically coded L-amino acids.
 13. The compound of claim 12, wherein X₈ is G, S, Y or R.
 14. The compound of claim 12, wherein said compound comprises a sequence of amino acids (SEQ ID NO:15): G G C T L R E W L H G G F C G G.
 15. The compound of claim 6, wherein said compound comprises a sequence of amino acids (SEQ ID NO:3):

    X.sub.8 G X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 W X.sub.7

where X₁ is L, M, P, Q, or V; X₂ is F, R, S, or T; X₃ is F, L, V, or W; X₄ is A, K, L, M, R, S, V, or T; X₅ is A, E, G, K, M, Q, R, S, or T; X₇ is C, I, K, L, M or V; and X₈ is any of the 20 genetically coded L-amino acids.
 16. The compound of claim 15, wherein X₁ is P; X₂ is T; X₃ is L; X₄ is R; X₅ is E or Q; X₇ is I or L (SEQ ID NO:4).
 17. The compound of claim 16, wherein said compound comprises a sequence of amino acids (SEQ ID NO:5):

    X.sub.9 X.sub.8 G X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 W X.sub.7

where X₈ is A, C, D, E, K, L, Q, R, S, T, or V; and X₉ is A, C, E, G, I, L, M, P, R, Q, S, T, or V.
 18. The compound of claim 17, wherein X₈ is D, E, or K; and X₉ is A or I.
 19. The compound of claim 18, wherein said compound is selected from the group consisting of (SEQ ID NOS: 6-13, respectively) G G C A D G P T L R E W I S F C G G; G N A D G P T L R Q W L E G R R P K N; G G C A D G P T L R E W I S F C G G K; T I K G P T L R Q W L K S R E H T S; S I E G P T L R E W L T S R T P H S; L A I E G P T L R Q W L H G N G R D T; C A D G P T L R E W I S F C; and I E G P T L R Q W L A A R A.
 20. The method of claim 4, wherein said compound that is administered to the patient comprises a sequence of amino acids (SEQ ID NO:15):

    C X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7

where X₂ is K, L, N, Q, R, S, T or V; X₃ is C, F, I, L, M, R, S or V; X₄ is any of the 20 genetically coded L-amino acids; X₅ is A, D, E, G, S, V or Y; X₆ is C, F, G, L, M, S, V, W or Y; and X₇ is C, G, I, K, L, M, N, R or V.
 21. The method of claim 20, wherein X₄ is A, E, G, H, K, L, M, P, Q, R, S, T or W.
 22. The method of claim 21, wherein X₂ is S or T; X₃ is L or R; X₄ is R; X₅ is D, E, or G; X₆ is F, L, or W; and X₇ is I, K, L, R, or V.
 23. The method of claim 22, wherein said compound that is administered to the patient comprises a sequence of amino acids (SEQ ID NO:15): G G C T L R E W L H G G F C G G.
 24. The method of claim 4, wherein the disorder susceptible to treatment with a thrombopoietin agonist is selected from the group consisting of:hematological disorders and thrombocytopenia resulting from chemotherapy, radiation therapy, or bone marrow transfusions.
 25. The method of claim 4, wherein said compound that is administered to the patient comprises a sequence of amino acids (SEQ ID NO:16):

    X.sub.8 G X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 W X.sub.7

where X₁ is L, M, P, Q, or V; X₂ is F, R, S, or T; X₃ is F, L, V, or W; X₄ is A, K, L, M, R, S, V, or T; X₅ is A, E, G, K, M, Q, R, S, or T; X₇ is C, I, K, L, M or V; and X₈ residue is any of the 20 genetically coded L-amino acids.
 26. The method of claim 25, wherein X₁ is P; X₂ is T; X₃ is L; X₄ is R; X₅ is E or Q; X₇ is I or L.
 27. The method of claim 26, wherein said compound comprises a sequence of amino acids (SEQ ID NO:5):

    X.sub.9 X.sub.8 G X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 W X.sub.7

where X₈ is A, C, D, E, K, L, Q, R, S, T, or V; and X₉ is A, C, E, G, I, L, M, P, R, Q, S, T, or V.
 28. The method of claim 27, wherein X₈ is D, E, or K; and X₉ is A or I.
 29. The method of claim 28, wherein the compound that is administered to the patient is selected from the group consisting of (SEQ ID NOS: 6-13, respectively) G G C A D G P T L R E W I S F C G G; G N A D G P T L R Q W L E G R R P K N; G G C A D G P T L R E W I S F C G G K; T I K G P T L R Q W L K S R E H T S; S I E G P T L R E W L T S R T P H S; L A I E G P T L R Q W L H G N G R D T; C A D G P T L R E W I S F C; and I E G P T L R Q W L A A R A.
 30. A compound that binds to thrombopoietin receptor, wherein said compound is selected from the group consisting of ##STR102##
 31. A method for treating a patient suffering from a disorder that is susceptible to treatment with a thrombopoietin agonist, comprising administering to the patient a compound selected from the group consisting of 